Bergenin exerts antitumor effects by promoting survivin degradation

doi:10.21203/rs.3.rs-1547129/v1

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Bergenin exerts antitumor effects by promoting survivin degradation

https://doi.org/10.21203/rs.3.rs-1547129/v1

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Survivin is overexpressed in multiple human malignancies and correlated with poor prognosis. In this study, we found that survivin is highly expressed in non-small cell lung cancer (NSCLC) tissues, and depletion of survivin inhibits tumorigenesis of NSCLC cells. Moreover, our results showed that bergenin, a natural product, exhibits antitumor activity against NSCLC cells. Bergenin suppresses cell viability and colony formation in A549, H1299 and H460 cells, and reduces the protein level of survivin dose-dependently. The mechanism study revealed that bergenin inhibits the phosphorylation of survivin by regulating Akt/Wee1/CDK1 signaling pathway, thereby enhancing the interaction between survivin and E3 ligase Fbxl7. In addition, bergenin enhances the expression of cleaved-PARP and -caspase 3, which promotes apoptosis of NSCLC cells. The in vivo study showed that the tumor volume, weight, and the expression of Ki67 and survivin were decreased obviously in bergenin treated group. Overall, our data indicate that bergenin has a significant antitumor activity and is a promising agent with therapeutic potential for NSCLC.

Non-small cell lung cancer

Survivin

Bergenin

ubiquitination

Non-small cell lung cancer (NSCLC) is the predominant subtype of lung cancer, accounting for 80–85%, and remains the primary cause of cancer-related mortality worldwide^1,2. NSCLC can be subdivided into adenocarcinoma (ADC), squamous cell carcinoma (SCC), large cell carcinoma (LCC), and mixed histology³. Recently, people have made growing efforts to improve detection methods to screen high-risk groups to achieve early diagnosis. However, only a small proportion of NSCLC patients (< 20%) are diagnosed at an early stage due to the lack of specific symptoms, and most of them (47%) are still diagnosed in the advanced stages⁴. The treatment methods of NSCLC include traditional surgery, radiotherapy/chemotherapy, targeted therapy, and immunotherapy^5,6. Although monotherapy or combination therapy has improved patients' quality of life and overall survival, patients are still at risk of tumor recurrence and progression. Continuing pre/clinical trials aimed to develop new drugs to improve the management of NSCLC is in urgent demand.

Survivin, a member of the inhibitors of apoptosis proteins (IAPs), plays a crucial role in regulating cell division and apoptosis. It is found in different subcellular localization (nucleus, cytoplasm, mitochondria and extracellular) with multiple functions. Nuclear survivin participates in the formation of the chromosome passenger complex (CPC) and regulates cell division, while cytoplasmic and mitochondrial expression is considered to function as apoptotic suppressor^7,8; the extracellular pool of survivin is secreted by tumor cells in the form of membrane vesicles and absorbed by surrounding cells, which is related to increase therapeutic resistance and promote tumor development⁹. Survivin is not observed in normal adult tissues but is highly expressed in embryonic tissues, cancer cells, and cancer stem cells such as lung cancer, bladder cancer, ovarian cancer, breast cancer and pancreatic ductal adenocarcinoma (PDAC), and this expression is associated with aggressiveness and progression of cancer^10–13. Recently, survivin has been served as a universal tumor-associated antigen and a biomarker of cancer progression. Therefore, targeting survivin is emerging as a promising therapeutic strategy in cancer.

Here, we investigated the antitumor effect of bergenin and revealed the underlying mechanism. The natural product bergenin may be a potential survivin inhibitor and an antitumor candidate for NSCLC therapy.

Cell culture and reagents

The immortalized lung epithelial cells NL20 and MRC5 and human NSCLC A549, H1299 and H460 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA). All cells were grown in a humidified incubator at 37℃ with 5% CO₂ and subjected to mycoplasma analysis routinely. The cell culture medium of RPMI-1640 and Fetal Bovine Serum (FBS) were obtained from Invitrogen (Grand Island, NY). The natural products bergenin, MG 132, and cycloheximide (CHX) were obtained from Selleck Chemicals (Houston, TX). Transfection reagent lipofectamine 2000 was purchased from Thermo Fisher Scientific (Waltham, MA). Antibodies against survivin, cleaved-caspase 3, cleaved-PARP, cleaved-caspase 3, cytochrome C, Bax, VDAC1, p-Survivin Thr34, p-Akt Ser473, p-Wee1 Ser642, p-CDK1 Tyr15 and p-CDK1 Thr161, ubiquitin, Flag-tag, HA-tag, α-Tubulin and β-actin were purchased from Cell Signaling Technology, Inc. (Beverly, MA). The anti-ki 67 and FbxL 7 antibodies were products of Abcam (Cambridge, United Kingdom).

MTS Assay.

The MTS assay, used to analyze cell viability, was performed as described previously¹⁴. Briefly, human NSCLC cells were seeded into the 96-well plates (2×10³ cells/well) and maintained overnight. Cells were incubated with various concentrations of bergenin for 24, 48, and 72 h. Then cell viability was analyzed according to the standard protocol after adding the MTS reagent (#G3580, Madison, WI) to the cell culture medium.

Anchorage-independent cell growth.

The Anchorage-independent cell growth assay was performed as described previously¹⁵. Briefly, Eagle’s basal medium consists of 0.6% agar, 10% FBS and various concentration of bergenin, which were loaded in a 6-well plate as an agar base. Human NSCLC cells were resuspended at a concentration of 8000 cells/ml and inoculated in 6-well plates containing 0.6% Basal Medium Eagle (0.3% agar, 10% FBS, and different doses of bergenin. Colonies were counted after maintaining for 2 weeks.

Clinical tissue sample collections.

A total of 39 cases of NSCLC tissues and matched adjacent non-tumor tissues were collected from 39 patients in the department of pathology, The Third Xiangya Hospital of Central South University, Changsha, Hunan, China. All the patients signed written informed consent and did not receive treatment before surgery.

Western blotting.

The cells were treated with bergenin and lysed in RIRA buffer (#89901, Thermo Fisher Scientific) at 4℃ for around 30min followed by centrifugation at 12,000 × rpm for 10min to collect supernatant as the whole-cell extract (WCE). Then the BCA protein assay kit (#23228, Thermo Fisher Scientific) was used to detect the WCE concentration. A total of 20ug WCE was subjected to SDS-PAGE gels and transferred to the PVDF membrane. Subsequently, the membranes are blocked with 5% non-fat milk for 1 h and incubated with primary and second antibodies. The ECL reagent (#34579, Thermo Fisher Scientific) was used to visualize the target protein expression.

Co-immunoprecipitation (Co-IP).

Total proteins from NSCLC cells cultured in 10 cm culture dishes were used for the Co-IP assay. Briefly, primary antibodies against HA or Flag were added to each cell lysate and incubated overnight at 4℃ with rotation. Protein A/G agarose beads were added and incubated with mild rotation at 4°C for 2 h to bind to the primary antibody. The agarose beads were washed with PBS and cell lysis buffer to wash off unbound antibodies. The agarose beads were resuspended in 20 µl 1x SDS-PAGE loading buffer and boiled and centrifuged to collect the supernatant for western blot analysis.

Generation of survivin knockdown stable cell lines.

To generate survivin knockdown stable cells, 293T cells were co-transfected with pLKO.1-shsurvivin lentivirus plasmids (TRCN0000073718, TRCN0000073721, Millipore Sigma), PSPAX2 and PMD2-G. The supernatant containing viral was collected at 72 h after transfection. NSCLS cells were grown at 70%-80% confluence and infected with the lentivirus and polybrene (5 µg/ml). The infected cells were cultured with fresh medium containing puromycin (1µg/ml) and maintained for 1 week for colony selection.

In vivo tumor growth.

The in vivo tumorigenesis was approved by the Institutional Animal Care and Use Committee (IACUC) of the Third Xiangya Hospital of Central South University (Changsha, China). A549 and H1299 cells (2×10⁶) were injected into the right flank of 6-week-old athymic nude mice (n = 5). Tumor growth was monitored, and volume was measured every 2 days. The tumor-bearing mice were randomly divided into two groups (n = 5) when tumor volume reached around 100 mm³. The treatment group was initiated bergenin (30 mg/kg) via intraperitoneal injection every two days, and the control group was administrated the vehicle control. The mice were euthanized with CO₂ (3 L/min) for 5 min at the endpoint. Tumor volume was calculated following the formula: length × width × width/2.

Immunohistochemical staining (IHC).

The IHC staining was performed as described previously¹⁶. Tumor tissues obtained from clinical samples and xenograft tumors were subjected to IHC analysis. Briefly, the tissue sections were deparaffinized and rehydrated. The antigen retrieval was performed subsequently by submerging into sodium citrate buffer (10 mM, pH 6.0) and boiled for 10 min. Then the tissue slides were treated with 3% H₂O₂ in methanol for 10 min, washed with PBS and blocked with 10% goat serum albumin, and incubated with primary antibodies overnight at 4℃. After hybridization with secondary antibodies at room temperature, the target protein was visualized using the DAB substrate and counterstained by hematoxylin.

Statistical analysis.

GraphPad Prism 5 (GraphPad 5.0, San Diego, CA, USA) software was used for statistical analysis. Data were performed from at least three independent determinations and represented as mean ± sd. Statistical comparisons between different groups were analyzed by Student's t-test or ANOVA. The Kaplan-Meier method was applied to estimate survival functions and the log-rank test was used to compare survival differences between groups. A probability value of p < 0.05 was used as the criterion for statistical significance (*P < 0.05, **P < 0.01, ***P < 0.001).

Survivin is overexpressed in NSCLC tumor tissues.

We first examined survivin protein expression in NSCLC tissues and adjacent tissues by immunohistochemistry (IHC). The result showed that survivin expression in tumor tissues is higher than in adjacent tissue (Fig. 1A). The Kaplan-Meier survival analyses revealed that the high level of survivin is correlated with worse overall survival (OS), first progression survival (FP), and post-progression survival (PPS) of NSCLC patients (Fig. 1B). To determine the oncogenic role of survivin, we generated survivin knockdown stable cell lines in A549 and H1299 cells. The MTS assay and the colony formation assay revealed that knockdown survivin inhibited cell viability and anchorage-independent cell growth significantly (Fig. 1C-D). We further conducted the athymic nude mouse xenograft models. We found that depletion of survivin robustly inhibited tumor growth. The tumor volume and weight were significantly reduced in the A549-shSurvivin xenografts (Fig. 1E-F). Consistently, the IHC data showed that the population of Ki67 positive cells was attenuated obviously in shsurvivin-expression xenograft tumors (Fig. 1G). These results indicated that survivin is overexpressed in NSCLC tissues and may play an important role in NSCLC tumorigenesis.

Bergenin inhibits the viability of NSCLC cells and exhibits little cytotoxicity.

Earlier studies have reported that natural plant extract bergenin (Fig. 2A) has several pharmacological activities, such as anti-inflammatory, hypolipidimic, antitumor and antimalarial, etc¹⁷. To determine the antitumor effect of bergenin on NSCLC cells, A549, H1299 and H460 cells were treated with different concentrations of bergenin. The MTS and colony formation data revealed that bergenin significantly inhibited the cell viability and colony formation dose-dependently (Fig. 2B-D). Exposure to bergenin (40 µM) for 48 h decreased the cell viability of all tested NSCLC cells by over 50%. Meanwhile, the anchorage-independent growth assay showed that the colony formation reduced over 75% of A549 and H1299 cells treated with 20 uM bergenin, and 40uM bergenin decreased the colony number by over 90% of all three cell lines. Moreover, to explore the effect of bergenin on non-tumor cells, the immortalized non-tumor lung tissue cells NL20 and MRC5 were treated with bergenin. The results showed that the cell viability did not decrease significantly even if the concentration of bergenin reached 160 uM (Fig. 2E). These results indicate that bergenin has antitumor effect against NSCLC cells and is well-tolerated in immortalized non-tumor cells.

Bergenin facilitates Fbxl7-mediated survivin ubiquitination and degradation in NSCLC cells.

To determine whether the antitumor effect of bergenin in NSCLC cells is through the regulation of survivin, we then detected the expression level of survivin in bergenin-treated NSCLC cells by immunoblotting (IB) assay. The data showed that survivin decreased in A549 and H1299 cells dose-dependently (Fig. 3A). Moreover, treatment with MG132, a proteasome inhibitor, restored the protein level of survivin in bergenin-treated 549 and H1299 cells (Fig. 3B-C). In addition, bergenin reduced the half-life of survivin substantiall (Fig. 3D), indicating that bergenin might affect the stability of survivin protein. We next used the co-IP analysis to investigate the ubiquitination of survivin in bergenin-treated cells. The result showed that bergenin promoted survivin ubiquitination obviously (Fig. 3E) and enhanced the interaction between survivin and the E3 ligase Fbxl7 (Fig. 3F). To determine whether E3 ligase Fbxl7 is indispensable for survivin ubiquitination in bergenin-treated NSCLC cells, we transfected siRNA to silence Fbxl7 in A549 cells and found that knockdown of Fbxl7 impaired bergenin-induced survivin ubiquitination (Fig. 3G). Furthermore, double mutation of the residues K90/91, two ubiquitination sites required for survivin degradation, compromised Fbxl7-induced survivin ubiquitination in A549 cells (Fig. 3H). Meanwhile, the survivin K90/91 mutant reduced bergenin-induced survivin ubiquitination significantly (Fig. 3I). These results imply that bergenin facilitates survivin ubiquitination and degradation by enhancing the interaction between survivin and Fbxl7.

Bergenin-inhibited Akt/Wee1/CDK1 pathway is required for survivin ubiquitination

Previous studies have shown that protein survivin phosphorylation on Thr34 plays an essential role in its biological function and stability. Therefore, we determined whether bergenin promotes survivin degradation via regulating its phosphorylation. As shown in Fig. 4A, bergenin suppressed survivin Thr34 phosphorylation dose-dependently in A549 and H1299 cells. Moreover, we found that after treating with bergenin, the phosphorylation of Akt (Ser473), the downstream target Wee1 (Ser642), CDK1 (Thr161) (a marker for CDK1 activation), and survivin (Thr34) is decreased dose-dependently. Consistently, CDK1 phosphorylation on Tyr15 is increased (Fig. 4B). These results indicate that Bergenin inhibited the Akt/Wee1/CDK1 signaling. This inhibitory effect was also observed in the experiment of knockdown Akt with siRNA (Fig. 4C). Moreover, transfection of Myr-Akt1, a constitutively activated Akt, compromised bergenin-induced reduction of phosphorylation of Wee1 (Ser642), CDK1 (Thr161), and survivin (Thr34) (Fig. 4D). Additionally, a series of experimental analysis data showed that overexpression of Myr-Akt1 rescued cell viability, increased the live cell population, down-regulated the activated cleaved-caspase 3 significantly, and decreased the protein level of cleaved-caspase 3 and -PAPR in bergenin-treated NSCLC cells (Fig. 4E-H). Compared with wild-type (WT) survivin, mutation of survivin Thr34 to Ala34 induced stronger ubiquitination of survivin in bergenin-treated A549 cells (Fig. 4I). Importantly, survivin WT, but not the T34A mutant, rescued cell viability (Fig. 4J) and increased the population of live cells (Fig. 4H). Collectively, our results demonstrate that inhibition of the Akt-Wee1-CDK1 signal pathway is required for survivin phosphorylation, which promotes survivin ubiquitination and degradation.

Bergenin promotes apoptosis in NSCLC cells.

To examine whether bergenin activates the apoptosis signaling in NSCLC cells, we tested the protein level of cleaved-PARP and -caspase 3 in bergenin-treated A549 and H1299 cells. The IB data showed that bergenin promoted the expression of cleaved-PARP and -caspase 3 dose-dependently (Fig. 5A). Consistently, the enzymatic activity of caspase 3 was significantly enhanced with bergenin treatment (Fig. 5B). Additionally, we examined changes in the intracellular location of apoptosis-related molecules and found that bergenin facilitated the release of cytochrome C from mitochondrial to the cytoplasm. With the increase of bergenin concentration, the protein Bax in the cytoplasm showed a decreasing trend, while that in mitochondria increased (Fig. 5C), suggesting that bergenin can activate the intrinsic apoptotic pathway. We next determined whether the destruction of survivin caused bergenin-induced apoptosis. The results showed that overexpression of survivin restored cell viability (Fig. 5D), compromised the activity of caspase 3 (Fig. 5E), and decreased the protein level of cleaved-caspase 3 and -PARP (Fig. 5F) in bergenin-treated A549 cells. Furthermore, subcellular fractions immunoblotting assay suggested that overexpression of survivin rescued bergenin-promoted intrinsic apoptosis, as the expression level of protein Bax and cytochrome C in cytoplasm and mitochondria were close to the control group without any treatment (Fig. 5G). Our data suggested that bergenin promotes mitochondrial apoptosis in NSCLC cells.

Butein inhibits in vivo tumor growth.

To verify the in vivo anti-tumor effect of bergenin, we performed xenograft mouse models using A549 and H1299 cells. Our data showed that treatment with bergenin significantly delayed the xenograft tumor growth. Compared with the vehicle control group, the tumor volume of the bergenin-treated group was reduced significantly (Fig. 6A-B), and tumor weight decreased by about two-thirds of A549 tumors (Fig. 6C). Consisitently, in the H1299 xenograft tumor model, bergenin exhibited similarly significant antitumor activity (Fig. 6D-6F). Similar to the in vitro findings, the IHC results revealed that bergenin decreased the population of Ki67 positive cells and the number of survivin positive cells in A549-derived tumors (Fig. 6G and 6H). Overall, our data implied that bergenin suppresses in vivo tumor development of NSCLC cells.

Accumulating evidence shows that survivin is upregulated in multiple human cancers, and high level of survivin is correlated with the proliferation, invasion, metastasis, recurrence, therapeutic resistance, and poor prognosis in most cancers¹⁸, including NSCLC^19–21. For instance, it’s found that survivin expression level was significantly higher in hepatocellular cancer (HCC) tissue than in normal tissue. And survivin overexpression was found to promot HepG2 cell proliferation, inhibit apotosis and induce invasive ability²². The data of a meta-analysis suggested that the level of survivin expression associated with the overall survival (OS) of NSCLC patients was significant and implied positive-survivin expression might be a prognostic factor for NSCLC patients²³. In the present study, we found that survivin is highly expressed in NSCLC tumor tissues and its expression level is negatively correlated with the prognosis of NSCLC patients. Knockout survivin by sgRNA plasmid transfection inhibited the cell proliferation, colony formation, and tumor growth in vivo. This evidence indicates that survivin plays a crucial role in the tumorigenesis of NSCLC.

Survivin regulation in human cancer cells is complicated and controlled at multiple levels, including transcription, translation, and posttranslational²⁴. Transcriptional regulation of survivin mRNA expression includes epigenetic modifications, transcription factors, signaling pathways, etc. Many transcription factors, such as SP1, NF-kB, GATA-1, STAT3, DEC1, and Notch, have emerged as important inducers of survivin expression in various tumors²⁵. The signaling pathways of Phosphoinositide 3-Kinase (PI3K)/Akt, Janus kinase (JAK)/signal transducer and activator of transcription (STAT) and Mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK), which are involved to tumor cellular proliferation, differentiation, migration and survival, exist a central role in regulating survivin expression under the action of different stimulators¹⁸. It has been found that modification of the phosphorylation sites (Thr34, Thr117, Ser20) is closely related to survivin's function. Protein kinase A (PKA) phosphorylates the serine 20 residue to suppress the anti-apoptotic function of survivin; cyclin-dependent kinase 1 (CDK1) phosphorylates the threonine 34, contributing to its stability and promoting interaction with the mitotic spindle^26–28. Post-translational modifications of ubiquitination and deubiquitination are important to regulate protein expression levels. At present, two E3 ligases (the F-box Protein Fbxl7 and the X-linked inhibitor of apoptosis (XIAP))^29,30 and some deubiquitinases (USP9X, CSN5, USP35)^31–33 have been reported to modulate the expression of survivin. According to the regulatory and functional mechanism of survivin, many drugs targeting survivin have been investigated in clinical and preclinical studies. For example, Piperine prevents the binding of survivin and Smac/DIABLO, making it available for inhibiting IAPs to inhibit the growth of human colon cancer³⁴. Tolfenamic acid (TA) downregulates survivin expression and suppresses pancreatic cancer development by inhibiting transcription factors SP1³⁵. Xanthohumol inhibits the tumorigenic properties of oral squamous cell carcinoma (OSCC) cells by decreasing survivin phosphorylation on Thr34 and facilitating E3 ligase Fbxl7-mediated ubiquitination, which in turn reduced survivin³⁶. In this study, we found that bergenin decreased survivin expression and reduced the protein half-life. Exploring the underlying mechanism revealed that bergenin inhibited survivin Thr34 phosphorylation by regulating the Akt/Wee1/CDK1 signal pathway, and enhanced the interaction between protein survivin and E3 ligase Fbxl7, which facilitates survivin ubiquitination and degradation. However, it is unclear whether bergenin also acts on other molecules of the IAPs family and other modification sites of survivin. Thus, further investigating the anti-tumor mechanism of bergenin will be meaningful.

Bergenin is reported to have several critical pharmacological activities, including hepatoprotective, anti-inflammatory, anti-oxidation, anti-diabetes, and antitumor effects^37–40. Previous studies showed that bergenin could attenuate bleomycin-induced pulmonary fibrosis via inhibiting the TGF-β1 signal⁴¹, relieve colitis in vivo through regulating PPARγ/SIRT1/NF-κB-p65 pathway⁴², protect acute lung injury by inhibiting NF-κB activation⁴³, and inhibit human bladder cancer progression via activating the PPARγ signal⁴⁴. In the current preclinical studies, the natural product bergenin is mainly used in non-neoplastic diseases, but its inhibitory effect on malignant tumors is rarely investigated. Our results found that bergenin promoted intrinsic apoptotic pathway by reducing survivin. In NSCLC cells, bergenin inhibited tumor cell proliferation, colony formation, and in vivo tumorigenesis by downregulating the Akt/Wee1/CDK1 signal pathway and reducing survivin stability. These findings indicate that bergenin could be a novel survivin inhibitor that deserves further study for NSCLC treatment.

In summary, we demonstrated that survivin was upregulated in NSCLC tumor tissues and required for maintaining malignant phenotypes of NSCLC cells. Bergenin reduced survivin expression dose-dependently and inhibited the phosphorylation of survivin by suppressing Akt/Wee1/CDK1 signal pathway, which enhanced the interaction between survivin and E3 ligase Fbxl7. This study suggests that bergenin appears to be an effective chemical targeting survivin and may serve as a promising therapeutic candidate for NSCLC.

Founding: This work was supported by the National Natural Science Foundation of China (No.81401548, 81972837, 82003203) and the Natural Science Foundation of Hunan Province (2021JJ31011, 2021JJ41058).

Competing interests: The authors have declared no conflicts of interest.

Author contributions:

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by [Xiaoying Li], [Yu Gan], [Shuangze Han], [Jinzhuang Liao], [Li Zhou] and [Wei Li]. Writing, review, and/or revision of the manuscript was written by [Xiaoying Li], [Li Zhou] and [Wei Li]. All authors read and approved the final manuscript.

Availability of data and materials

Materials are available upon request.

Ethics approval and consent to participate.

The animal experiments were approved by the Medical Research Animal Ethics Committee, Central South University, China.

Consent for publication

Not applicable.

Tan S et al (2018) Circular RNA F-circEA produced from EML4-ALK fusion gene as a novel liquid biopsy biomarker for non-small cell lung cancer. Cell Res 28:693–695. doi:10.1038/s41422-018-0033-7
Bray F et al (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68:394–424. doi:10.3322/caac.21492
Travis WD et al (2015) The 2015 World Health Organization Classification of Lung Tumors: Impact of Genetic, Clinical and Radiologic Advances Since the 2004 Classification. J Thorac Oncol 10:1243–1260. doi:10.1097/JTO.0000000000000630
Šutić M et al (2021) Diagnostic, Predictive, and Prognostic Biomarkers in Non-Small Cell Lung Cancer (NSCLC) Management. J Pers Med 11. doi:10.3390/jpm11111102
Duma N, Santana-Davila R, Molina JR (2019) Non-Small Cell Lung Cancer: Epidemiology, Screening, Diagnosis, and Treatment. Mayo Clin Proc 94:1623–1640. doi:10.1016/j.mayocp.2019.01.013
Mamdani H, Matosevic S, Khalid AB, Durm G, Jalal S (2022) I. Immunotherapy in Lung Cancer: Current Landscape and Future Directions. Front Immunol 13:823618. doi:10.3389/fimmu.2022.823618
Dohi T, Beltrami E, Wall NR, Plescia J, Altieri DC (2004) Mitochondrial survivin inhibits apoptosis and promotes tumorigenesis. J Clin Invest 114:1117–1127
Li F, Yang J, Ramnath N, Javle MM, Tan D (2005) Nuclear or cytoplasmic expression of survivin: what is the significance? Int J Cancer 114:509–512
Khan S, Bennit HF, Wall NR (2015) The emerging role of exosomes in survivin secretion. Histol Histopathol 30:43–50. doi:10.14670/HH-30.43
Zhou L et al (2018) High nuclear Survivin expression as a poor prognostic marker in pancreatic ductal adenocarcinoma. J Surg Oncol 118:1115–1121. doi:10.1002/jso.25253
Arista-Romero M, Cascante A, Fornaguera C, Borrós S (2021) Role of Survivin in Bladder Cancer: Issues to Be Overcome When Designing an Efficient Dual Nano-Therapy. Pharmaceutics 13. doi:10.3390/pharmaceutics13111959
Puskas R et al (2021) Circulating Survivin Protein Levels in Lung Cancer Patients Treated With Platinum-Based Chemotherapy. Pathol Oncol Res 27:631969. doi:10.3389/pore.2021.631969
Nan XW et al (2019) Survivin Promotes Piperlongumine Resistance in Ovarian Cancer. Front Oncol 9:1345. doi:10.3389/fonc.2019.01345
Li W et al (2018) Repression of Noxa by Bmi1 contributes to deguelin-induced apoptosis in non-small cell lung cancer cells. J Cell Mol Med 22:6213–6227. doi:10.1111/jcmm.13908
Li M et al (2021) Targeting Aurora B kinase with Tanshinone IIA suppresses tumor growth and overcomes radioresistance. Cell Death Dis 12:152. doi:10.1038/s41419-021-03434-z
Yu X et al (2019) Skp2-mediated ubiquitination and mitochondrial localization of Akt drive tumor growth and chemoresistance to cisplatin. Oncogene 38:7457–7472. doi:10.1038/s41388-019-0955-7
Gao XJ et al (2015) Bergenin Plays an Anti-Inflammatory Role via the Modulation of MAPK and NF-κB Signaling Pathways in a Mouse Model of LPS-Induced Mastitis. Inflammation 38:1142–1150. doi:10.1007/s10753-014-0079-8
Martínez-García D, Manero-Rupérez N, Quesada R, Korrodi-Gregório L (2019) Soto-Cerrato, V. Therapeutic strategies involving survivin inhibition in cancer. Med Res Rev 39:887–909. doi:10.1002/med.21547
Zhang LQ et al (2012) Prognostic value of survivin in patients with non-small cell lung carcinoma: a systematic review with meta-analysis. PLoS ONE 7:e34100. doi:10.1371/journal.pone.0034100
Karczmarek-Borowska B et al (2005) Survivin antiapoptotic gene expression as a prognostic factor in non-small cell lung cancer: in situ hybridization study. Folia Histochem Cytobiol 43:237–242
Jaiswal PK, Goel A, Mittal RD, Survivin (2015) A molecular biomarker in cancer. Indian J Med Res 141:389–397. doi:10.4103/0971-5916.159250
Yu J, Wang Z, Zhang H, Wang Y, Li DQ (2021) Survivin-positive circulating tumor cells as a marker for metastasis of hepatocellular carcinoma. World J Gastroenterol 27:7546–7562. doi:10.3748/wjg.v27.i43.7546
Fan J, Wang L, Jiang G-N, He W-X, Ding J-A (2008) The role of survivin on overall survival of non-small cell lung cancer, a meta-analysis of published literatures. Lung Cancer 61:91–96. doi:10.1016/j.lungcan.2007.11.011
Li F, Aljahdali I, Ling X (2019) Cancer therapeutics using survivin BIRC5 as a target: what can we do after over two decades of study? J Exp Clin Cancer Res 38:368. doi:10.1186/s13046-019-1362-1
Rafatmanesh A et al (2020) The survivin molecule as a double-edged sword in cellular physiologic and pathologic conditions and its role as a potential biomarker and therapeutic target in cancer. J Cell Physiol 235:725–744. doi:10.1002/jcp.29027
Wheatley SP, Henzing AJ, Dodson H, Khaled W, Earnshaw WC (2004) Aurora-B phosphorylation in vitro identifies a residue of survivin that is essential for its localization and binding to inner centromere protein (INCENP) in vivo. J Biol Chem 279:5655–5660
Dohi T, Xia F, Altieri DC (2007) Compartmentalized phosphorylation of IAP by protein kinase A regulates cytoprotection. Mol Cell 27:17–28
O'Connor DS et al (2000) Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. Proc Natl Acad Sci U S A 97:13103–13107
Liu Y et al (2015) The Proapoptotic F-box Protein Fbxl7 Regulates Mitochondrial Function by Mediating the Ubiquitylation and Proteasomal Degradation of Survivin. J Biol Chem 290:11843–11852. doi:10.1074/jbc.M114.629931
Arora V et al (2007) Degradation of survivin by the X-linked inhibitor of apoptosis (XIAP)-XAF1 complex. J Biol Chem 282:26202–26209
Chen H, Yang F, Li X, Gong Z-J, Wang L-W (2018) Long noncoding RNA LNC473 inhibits the ubiquitination of survivin via association with USP9X and enhances cell proliferation and invasion in hepatocellular carcinoma cells. Biochem Biophys Res Commun 499:702–710. doi:10.1016/j.bbrc.2018.03.215
Li J, Li Y, Wang B, Ma Y, Chen P (2018) CSN5/Jab1 facilitates non-small cell lung cancer cell growth through stabilizing survivin. Biochem Biophys Res Commun 500:132–138. doi:10.1016/j.bbrc.2018.03.183
Wang W et al (2021) Regulation of survivin protein stability by USP35 is evolutionarily conserved. Biochem Biophys Res Commun 574:48–55. doi:10.1016/j.bbrc.2021.08.050
Yaffe PB, Coombs P, Doucette MR, Walsh CD, Hoskin DW (2015) Piperine, an alkaloid from black pepper, inhibits growth of human colon cancer cells via G1 arrest and apoptosis triggered by endoplasmic reticulum stress. Mol Carcinog 54:1070–1085. doi:10.1002/mc.22176
Hurtado M et al (2018) Novel Survivin Inhibitor for Suppressing Pancreatic Cancer Cells Growth via Downregulating Sp1 and Sp3 Transcription Factors. Cell Physiol Biochem 51:1894–1907. doi:10.1159/000495715
Li M et al (2020) Promotion of ubiquitination-dependent survivin destruction contributes to xanthohumol-mediated tumor suppression and overcomes radioresistance in human oral squamous cell carcinoma. J Experimental Clin Cancer Research: CR 39:88. doi:10.1186/s13046-020-01593-z
Zhang G et al (2021) Bergenin alleviates H(2) O(2) -induced oxidative stress and apoptosis in nucleus pulposus cells: Involvement of the PPAR-γ/NF-κB pathway. Environ Toxicol 36:2541–2550. doi:10.1002/tox.23368
Rajput SA, Mirza MR, Choudhary MI (2020) Bergenin protects pancreatic beta cells against cytokine-induced apoptosis in INS-1E cells. PLoS ONE 15:e0241349. doi:10.1371/journal.pone.0241349
Xiang S, Chen K, Xu L, Wang T, Guo C (2020) Bergenin Exerts Hepatoprotective Effects by Inhibiting the Release of Inflammatory Factors, Apoptosis and Autophagy via the PPAR-γ Pathway. Drug Des Devel Ther 14:129–143. doi:10.2147/dddt.S229063
Zhang J et al (2013) Cancer chemopreventive effect of bergenin from Peltophorum pterocarpum wood. Chem Biodivers 10:1866–1875. doi:10.1002/cbdv.201300182
Li X et al (2021) Bergenin attenuates bleomycin-induced pulmonary fibrosis in mice via inhibiting TGF-β1 signaling pathway. Phytother Res 35:5808–5822. doi:10.1002/ptr.7239
Wang K et al (2017) Acting as an Agonist of PPARγ, Ameliorates Experimental Colitis in Mice through Improving Expression of SIRT1, and Therefore Inhibiting NF-κB-Mediated Macrophage Activation. Front Pharmacol 8:981. doi:10.3389/fphar.2017.00981. Bergenin
Yang S et al (2017) The natural product bergenin ameliorates lipopolysaccharide-induced acute lung injury by inhibiting NF-kappaB activition. J Ethnopharmacol 200:147–155. doi:10.1016/j.jep.2017.02.013
Liu J et al (2021) Bergenin inhibits bladder cancer progression via activating the PPARγ/PTEN/Akt signal pathway. Drug Dev Res 82:278–286. doi:10.1002/ddr.21751

No competing interests reported.

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Bergenin exerts antitumor effects by promoting survivin degradation

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Abstract

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Introduction

Materials And Methods

Cell culture and reagents

Statistical analysis.

Results

Bergenin-inhibited Akt/Wee1/CDK1 pathway is required for survivin ubiquitination

Discussion

Conclusion

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References

Additional Declarations

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