H. pylori infection promotes the proliferation of gastric cancer via CDK1 expression

doi:10.21203/rs.3.rs-3247316/v1

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H. pylori infection promotes the proliferation of gastric cancer via CDK1 expression

https://doi.org/10.21203/rs.3.rs-3247316/v1

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Backgroud: The pervasiveness of H. pylori infection contributes to numerous gastrointestinal disorders, thus posing a significant challenge for patients and society alike. This research examines the influence of CDK1 in H. pylori infected gastric cancer.

Methods: The presence of CDK1 in H. pylori infected GC tissues and cell lines was ascertained through immunohistochemical fluorescence staining, Real-time PCR, and western blot methodologies. Moreover, CDK1 was silenced in GC cells via siRNA, and the mobility and invasive capacity of GC cells with CDK1 suppression were evaluated using Transwell assays. In the end, the levels of inflammation and cell apoptosis were assessed by ELISA and flow cytometry.

Results: The findings suggest that CDK1 is elevated in GC tissues and cell lines, showing a significant rise in H. pylori infected GC. Concurrently, H. pylori infection enhances the migratory and invasive potential by modulating CDK1. Further, H. pylori incites a robust cellular inflammatory response and apoptosis by modulating CDK1.

Conclusion: The study concludes that H. pylori infection can alter multiple physiological processes in host cells by controlling CDK1, implying that CDK1 could serve as a potential molecular target for combating H. pylori infection.

gastric cancer

H. pylori

CDK1

inflammatory response

apoptosis

H. pylori is a gram-negative, spiral-shaped, microaerophilic bacterium that colonizes the gastric epithelium in a manner that results in the most prevalent bacterial infection worldwide [1]. Despite the fact that the majority of H. pylori carriers are asymptomatic, all will eventually develop gastritis, which could progress to gastroduodenal ulcers, gastric carcinoma, and lymphoma of the associated lymphoid tissue of the mucosa over time [2]. Approximately 5.5% of all cancers worldwide may be traced back to infections, and H. pylori is now considered the most common cause of these diseases [3]. Peptic ulcer illness occurs in 10% of infected people, gastric cancer (GC) develops in 1–3%, and mucosa-associated lymphoid tissue (MALT) lymphoma occurs in 0.1% [4, 5]. Although a substantial proportion of H. pylori-infected individuals exhibit no symptoms throughout their lives, they unfailingly develop chronic inflammation [6]. Infection with H. pylori is the most prevalent risk factor for gastric cancers, with adenocarcinoma being the most prevalent type [7]. Recent studies [8, 9] have revealed that nearly half of the Asian population, particularly in East Asia, is infected with H. pylori. Therefore, elucidating the molecular mechanism of H. pylori-induced carcinogenesis is essential for the development of new anti-GC strategies.

CDK1 is a member of the cyclin-dependent kinase (CDK) family of serine/threonine kinases that plays a crucial role in cell cycle progression [10]. When it comes to facilitating the G2-M transition and regulating the G1 progression and G1-S transition, CDK1 is the only necessary CDK [11]. CDK1 has been shown to have a role in DNA repair through homologous recombination and checkpoint activation [12, 13], two hallmarks of the DNA damage response (DDR). Recent studies have demonstrated that CDK1 has a major impact on cancer cell proliferation [14], in addition to its role in regulating the cell cycle. Epithelial ovarian cancer has been associated to the buildup of cytoplasmic CDK1 and poor prognosis [15]. Moreover, CDK1 overexpression is frequently observed in hepatocellular carcinoma and has been linked to tumor progression [16]. Zhu et al. found that the increase of CDK1 expression mediated by H. pylori-driven NF-κB activation is dependent on -catenin signaling, strongly indicating that the oncogenic functions of CDK1 may contribute to H. pylori-induced gastric tumorigenesis [17].

In this study, we intend to investigate the regulatory mechanisms of CDK1 in H. pylori-infected gastric cancer in order to elucidate their molecular relationships and regulatory mechanisms.

Tissue samples

GC tissues were obtained from patients undergoing surgical resection of GC. All gastric cancer tissues undergo urease test to screen out H. pylori positive tissues. The Ethics Committee of Chongqing University Cancer Hospital approve the protocols (CQUCH2019021) for acquiring all tissue samples and obtained the patient's informed consent form.

H. pylori culture

H. pylori strains were acquired from the Chinese Center for Disease Control and Prevention and cultivated for one week at 37°C and 5% O₂ on commercial H. pylori culture plates.

Cell culture

Gastric epithelial cells (GES-1) and GC cells (MGC-803) were acquired from the Cell Center of Shanghai Chinese Academy of Sciences. GES-1 and MGC-803 cells were cultured in DMEM with 10% fetal bovine serum at the environment of 37°C with 5% CO₂. The cells were treated with H. pylori in DMEM without fetal bovine serum and antibiotics at a multiplicity of infection (MOI) of 100:1. The co-culture was maintained for 48 hours at 37°C and 5% CO₂.

Cell transfection

The CDK1 siRNA and negative control sequences were designed and synthesized in accordance with reference [18] for cell transfection. MGC-803 cells were classified as control (Con), negative control (NC), and siCDK1 groups. Three times 105 cells per well were inoculated in a 12-well plate and incubated at 37°C and 5% CO₂ for 12 hours. Prior to transfection, 5 µl of siRNA was diluted in 200 µl of serum-free Opti-MEM (no.cat. 51985091, Gibco; Thermo Fisher Scientific, Inc., USA). The samples were incubated at 37°C for 30 minutes after the addition of Lipofectamine TM 3000 (no.cat. L3000001, Thermo Fisher Scientific, Inc., USA) to the diluted siRNA solution (total volume 400 l). Each well received 200 µl of this solution, and the cell culture plate was shaken gently to mix the culture solution. Cells that had been transfected were then cultured for further experiments.

QRT-PCR assay

Total RNA was isolated from cells using Trizol reagent (no.cat. 15596026, Thermo Fisher Scientific, Inc., USA) according to the manufacturer's instructions. Using the PrimeScript RT Reagent Kit (RR037A, TaKaRa, Inc., Dalian, China), 0.5 mg of total RNA was reverse-transcribed into cDNA. RT-PCR was performed on an Eppendorf Mastercycler PCR using SYBR Green qPCR Master Mix (no.cat. HY-K0501A, TaKaRa Inc., Dalian, China) according to manufacturer's instructions. After 5 minutes of initial denaturation at 94°C, the sample underwent 38 cycles of denaturation for 30 s, annealing for 25 s, and extension for 25 s at a temperature of 72°C. The following primer sequences were devised and synthesized by Invitrogen Company for RT-PCR: CDK1, F 5'-CTGGCTGATTTTGGCCTTGC-3' and R 5'-GAGTAACGAGCTGACC

CCAG; GAPDH, F 5'-ATGGATCCTTATGCGCGAAATTCTTC-3' and R 5'-ACACATCATATTCTTGGCATCCCAC-3'. GAPDH served as a quality control measure. The relative gene expression was determined with the 2^−ΔΔCt method [19].

Assays of migration and invasion

Using a Transwell apparatus (no.cat. CLS3460, Corning, Inc., New York, USA) with Matrigel (no.cat. E6909, Thermo Fisher Scientific, Inc., USA) in the upper chamber, invasion capability was evaluated. Without Matrigel, the same procedure was followed for the migration test. Sorted into NC, siCDK1, and siCDK1-Hp groups, transfected MGC-803 cells were cultured in DMEM medium without FBS for 12 hours. The upper compartment was inoculated with 5×10⁴ cells in 0.3 ml of DMEM medium without FBS, while the lower compartment was supplemented with DMEM medium containing 20% FBS. After 24 hours of incubation, the cells on the upper surface were removed, and the migrated cells on the lower surface were fixed and stained at room temperature for 15 minutes with 0.1% g/ml crystal violet (no.cat. C0121, Beyotime Biotechnology, Inc., China). In each experiment, five random fields were analyzed with an Olympus IX71 Imaging System (Olympus, Inc.; magnification, ×100) to ascertain the number of cells that had completely invaded the filter. Each experiment was conducted a minimum of three times and evaluated in duplicate.

Western blot analysis

The cells were lysed with RIPA lysis solution (no.cat. P0013C, Beyotime Biotechnology, China) and the supernatants were collected for Western blot analysis. Using a BCA reagent (no. cat. P0012S, Beyotime Biotechnology, China), the protein concentrations were determined. On 10% SDS-PAGE, 20 g of proteins per lane were separated and transferred to nitrocellulose membranes. Blots were blocked with 5% nonfat milk at 25°C for 1 hour, followed by an overnight incubation at 4°C with primary monoclonal antibodies specific for CDK1, P53, P27, CyclinA2, -Tubulin, and GAPDH (ProteinTech Group, China). After development, each blot was rinsed three times in 1X TBST (catalog number ST677, Beyotime Biotechnology, China). The blots were then visualized with a ChemiScope 6200 Touch Integrated Chemiluminescence Imaging System (Shanghai Qinxiang Scientific Instrument Co., China). ImageJ version 2.1.4.7 was used to analyze band intensity.

Inflammatory factors analysis

Inflammatory indicators were analyzed by measuring the amounts of IL-8 and TNF- in supernatants from the NC, siCDK1, and siCDK1-Hp groups using enzyme-linked immunosorbent assay (ELISA) (cat no. KE00006 and KE00154, ProteinTech Group, China) per the manufacturer's instructions. The levels of IL-8 and TNF- were calculated with the help of standard curves and then expressed as picograms per microliter. At least triplicates of each experiment were performed.

Apoptosis analysis

Flow cytometry with the Annexin V-FITC/PI apoptosis reagent (co.no. E-CK-A211, Elabscience, China) was used to analyze apoptosis in the NC, siCDK1, and siCDK1-Hp cell groups, according to the manufacturer's instructions. The cells were rinsed three times with PBS, resuspended in 1X binding buffer, stained with annexin V FITC and propidium iodide (PI), and incubated for 15 minutes at 25°C in the dark. The apoptotic rate of the cells was determined using a flow cytometer (BD Biosciences, USA). Each experiment was replicated a minimum of three times.

Immunohistochemistry analysis

6 GC tissues, 5 H. pylori-infected GC tissues, and 6 paracancerous tissues (Shanghai Xinchao Biotechnology, China) were processed overnight at room temperature with 1X TBST (no.cat. ST677, Beyotime Biotechnology, China) for immunohistochemical staining. Next, a primary monoclonal antibody specific for CDK1 (1:24,000; cat. no. 67575-1-Ig, ProteinTech Group, Inc., China) was incubated with the tissues overnight at 4°C. Tissues were washed three times with phosphate-buffered saline (no.cat. C0221A, Beyotime Biotechnology, China) after development. The tissues were then treated with a secondary antibody, goat anti-mouse IgG (H + L) FITC-labeled (cat. no. A0568, Beyotime Biotechnology, China), for two hours at 25°C. Images were taken using a Shanghai Qinxiang Scientific Instrument Co., China's ChemiScope 6200 Touch Integrated Chemiluminescence Imaging System.

Statistical analysis

For the purposes of statistical analysis and graph plotting, GraphPad Prism 9.0 (GraphPad Software, Inc. A one-way analysis of variance followed by Tukey's post hoc test and the Student's t-test were used for statistical analysis. The data are shown as Mean Standard Error of Mean with a significance level of P 0.05.

Analysis of CDK1 in GC tissues

Immunohistochemical fluorescence staining analysis revealed that CDK1 protein expression was substantially elevated in GC tissues compared to adjacent normal tissues (Fig. 1A, 1B). Notably, CDK1 proteins exhibited a significant increase in GC tissues infected with H. pylori compared to normal and GC tissues, indicating that H. pylori infection may increase CDK1 expression (Fig. 1A, 1B).

H. pylori infection influences CDK1 expression

In order to investigate the association between CDK1 and H. pylori infection further, the expression of CDK1 at protein and gene levels in GES-1 and MGC-803 cells infected with H. pylori was measured through qRT-PCR and western blotting. These results indicated that H. pylori may stimulate CDK1 expression in GES-1 and MGC-803 cells (Fig. 2A-2F). In addition, CDK1 was found to be overexpressed in MGC-803 cells compared to GES-1 cells, both in the presence and absence of H. pylori infection (Fig. 2G, 2H). These findings suggest that H. pylori infection may impact CDK1 expression.

H. pylori regulates CDK1 expression confirmed by siRNA knockdown

In order to confirm that H. pylori infection could regulate CDK1 expression, the effect of H. pylori on CDK1 was examined by inhibiting CDK1 expression. Using siRNA, the expression of CDK1 in MGC-803 cells was effectively suppressed (Fig. 3A-3C). Additionally, it was found that infection with H. pylori substantially increased CDK1 expression in CDK1- siRNA knockdown cells (Fig. 3D-3F). These experiments with siRNA inhibition demonstrated that H. pylori is capable of regulating CDK1 expression.

H. pylori regulates the migration and invasion of GC cells through CDK1

Previous research, such as that of Song et al. has shown that H. pylori can facilitate the migration and invasion of GC cells [20]. CDK1 is necessary for cell proliferation and invasion in addition to its role in regulating the cell cycle. In our study, CDK1 decrease inhibited the migration and invasion abilities of GC cells, while H. pylori infection promoted these processes through CDK1 overexpression (Fig. 4A-4D). These findings suggest that H. pylori may influence the proliferation of GC cells via CDK1.

The inflammation induced by H. pylori through CDK1

CDK1 has been identified as a signaling intermediary that links infection and inflammatory signaling to the activation of β-catenin, a key transcription network co-activator in GC [17]. The levels of the inflammatory factors IL-8 and TNF-α were measured in the supernatants of H. pylori-infected CDK1 knockdown GC cells. The results demonstrated that CDK1 reduction inhibited the expression of IL-8 and TNF-α in GC cells, whereas H. pylori infection increased the expression of these inflammatory factors (Fig. 5A-5B). These findings imply that infection with H. pylori could induce inflammatory responses by regulating CDK1.

The apoptosis of GC cell regulated by H. pylori through CDK1

CDK1 primarily regulates cells to enter the G2 phase from the M phase and influences cell proliferation. In this study, CDK1 knockdown substantially increased the apoptosis of MGC-803 cells, and H. pylori infection further intensified cell apoptosis in these CDK1 knockdown cells by upregulating CDK1 (Fig. 6A-6B).

Analysis of cell cycle proteins

CDK1 not only coordinates and promotes cell cycle processes during mitosis as a checkpoint, but also participates in the p53 signaling pathway. The expression levels of the cell cycle-related proteins P53, P27, and CyclinA2 were determined using western blot. Results showed that the knockdown of CDK1 increased the expression of P53 and P27 significantly, but the infection of H. pylori suppressed their expression in CDK1 knocked down cells (Fig. 7A-7B). In addition, the expression of CyclinA2 decreased in CDK1 knocked down cells obviously, but H. pylori infection also inhibited the expression of CyclinA2 (Fig. 7C-7D).

H. pylori is classified as a class I carcinogen by the World Health Organization. Chronic H. pylori infection and inflammation are the most prominent etiologic factors in the development of human gastric cancer [21, 22]. The G2/M phase transition and the activation of the homologous recombination (HR) DNA repair pathway are required for the progression of the cell cycle [23]. Zhu and colleagues discovered an increase in CDK1 expression as a result of H. pylori-mediated NF-κB activation, indicating that CDK1's oncogenic functions may contribute to H. pylori-induced gastric tumorigenesis [17]. This study aimed to determine the functions of CDK1 in H. pylori-infected GC and the association between abnormal expression and H. pylori infection.

CDK1 was substantially overexpressed in GC tissues, and its expression was twofold in GC tissues infected with H. pylori. Reduced CDK1 expression could inhibit GC cell proliferation and metastasis [24]. Therefore, we utilized siRNA technology to effectively suppress CDK1 expression in MGC-803 cells in vitro. However, infection with H. pylori increased CDK1 expression in these cells. Moreover, our findings indicate that low CDK1 expression inhibits the migration and invasion capabilities of gastric cancer (GC) cells, whereas H. pylori infection promotes these capabilities by increasing CDK1 expression, as demonstrated by Transwell experiments.

Infection with H. pylori stimulates immune and non-immune cells to generate a variety of inflammatory mediators, including IL-6, IL-8, TNF-α and MMPs, thereby maintaining mucosal inflammation and degradation [25, 26]. Zhu et al. discovered that CDK1-mediated activation of β-catenin signaling promotes cancer cell proliferation and illustrates a mechanistic link between infection, inflammation, and gastric malignancy in which CDK1 plays a central role [17]. In our study, we discovered that CDK1 downregulation inhibited the inflammatory response, for example, by diminishing the production of IL-8 and TNF-α. H. pylori infection, on the other hand, reactivated the inflammatory response by increasing CDK1 expression.

In vivo, H. pylori is known to stimulate both proliferation and apoptosis of epithelial cells; however, the H. pylori-cagA genotype induces more proliferation than apoptosis [27]. However, the molecular regulatory mechanism of proliferation and apoptosis induced by H. pylori infection is still unknown. Nie et al. demonstrated that CDK1 downregulation inhibited neuronal apoptosis and autophagy, thereby facilitating the recovery of motor function following spinal cord injury [28]. CDK1 suppression inhibited apoptosis in MGC-803 cells, whereas H. pylori infection increased apoptosis by promoting CDK1. CDK1 may also interact indirectly with the tumor suppressor protein P53 and the apoptosis regulator CyclinA2 to regulate the cell cycle and apoptosis [29, 30]. P27 is an endogenous CDK inhibitor and negatively modulates CDK1 expression to suppress the proliferation of cancer cells [31]. We discovered that lower CDK1 expression promoted the expression of P53 and P27, but inhibited CyclinA2 expression. In contrast, H. pylori infection functions the opposite effect on CDK1 knockdown GC cells.

CDK1 regulates the proliferation and apoptosis of H. pylori-infected GC cells, indicating that it is a potential prognostic and therapeutic target for treating H. pylori infection. However, the specific molecular mechanisms of CDK1 in H. pylori infection require additional studies in the future.

Ethics approval and consent to participate

The Ethics Committee of Chongqing University Cancer Hospital approved the protocols (CQUCH2019021). Informed consent was obtained from all participants involved in the study. All procedures in this research were performed in accordance with the Declaration of Helsinki and relevant regulations.

Consent for publication

Not applicable.

Availability of data and materials

Clinical data cannot be published due to ethical consideration. Please contact the corresponding author for potential access of clinical data.

Competing interests

The authors declare that they have no competing interests.

Funding

None

Authors' contributions

SJ and ZZ contributed to the conception or design of the work, and did the experiments. YX completed the data analysis and revised this manuscript. All authors read and approved the final manuscript.

Acknowledgements

Not applicable.

Hooi JKY, Lai WY, Ng WK, Suen MMY, Underwood FE, Tanyingoh D, et al. Global Prevalence of Helicobacter pylori Infection: Systematic Review and Meta-Analysis. Gastroenterology. 2017; 153: 420-429.
Alipour M. Molecular Mechanism of Helicobacter pylori-Induced Gastric Cancer. J Gastrointest Cancer. 2021; 52: 23-30.
Correa P. Gastric cancer: overview. Gastroenterol Clin North Am. 2013; 42: 211-217.
Wroblewski LE, Peek RM Jr. Helicobacter pylori, Cancer, and the Gastric Microbiota. Adv Exp Med Biol. 2016; 908:393-408.
Stewart OA, Wu F, Chen Y. The role of gastric microbiota in gastric cancer. Gut Microbes. 2020; 11: 1220-1230.
Engstrand L, Graham DY. Microbiome and Gastric Cancer. Dig Dis Sci. 2020; 65: 865-873.
Bornschein J, Malfertheiner P. Helicobacter pylori and gastric cancer. Dig Dis. 2014; 32: 249-64.
Choi YJ, Kim N. Gastric cancer and family history. Korean J Intern Med. 2016; 31: 1042-1053.
Hussein NR. Helicobacter pylori and gastric cancer in the Middle East: a new enigma? World J Gastroenterol. 2010; 16: 3226-3234.
Diril MK, Ratnacaram CK, Padmakumar VC, Du T, Wasser M, Coppola V, et al. Cyclin-dependent kinase 1 (Cdk1) is essential for cell division and suppression of DNA re-replication but not for liver regeneration. Proc. Natl. Acad. Sci. USA. 2012; 109: 3826-3831.
Haneke K, Schott J, Lindner D, Hollensen AK, Damgaard CK, Mongis C, et al. CDK1 couples proliferation with protein synthesis. J Cell Biol. 2020; 219: e201906147.
Enserink JM, Kolodner RD. An overview of Cdk1-controlled targets and processes. Cell Div. 2010; 5: 11.
Sunada S, Saito H, Zhang D, Xu Z, Miki Y. CDK1 inhibitor controls G2/M phase transition and reverses DNA damage sensitivity. Biochem Biophys Res Commun. 2021; 550: 56-61.
Dai Y, Hu S, Bai S, Li J, Yang N, Zhai P, et al. CDK1 promotes the proliferation of melanocytes in Rex rabbits. Genes Genomics. 2022; 44: 1191-1199.
Yang W, Cho H, Shin HY, Chung JY, Kang ES, Lee EJ, et al. Accumulation of cytoplasmic Cdk1 is associated with cancer growth and survival rate in epithelial ovarian cancer. Oncotarget. 2016; 7(31): 49481-49497.
Wu CX, Wang XQ, Chok SH, Man K, Tsang SHY, Chan ACY, et al. Blocking CDK1/PDK1/β-Catenin signaling by CDK1 inhibitor RO3306 increased the efficacy of sorafenib treatment by targeting cancer stem cells in a preclinical model of hepatocellular carcinoma. Theranostics. 2018; 8: 3737-3750.
Zhu S, Al-Mathkour M, Cao L, Khalafi S, Chen Z, Poveda J, et al. CDK1 bridges NF-κB and β-catenin signaling in response to H. pylori infection in gastric tumorigenesis. Cell Rep. 2023; 42: 112005.
Liu Y, Zhu YH, Mao CQ, Dou S, Shen S, Tan ZB, et al. Triple negative breast cancer therapy with CDK1 siRNA delivered by cationic lipid assisted PEG-PLA nanoparticles. J Control Release. 2014; 192:114-121.
Damgaard MV, Treebak JT. Protocol for qPCR analysis that corrects for cDNA amplification efficiency. STAR Protoc. 2022; 3: 101515.
Song Y, Liu G, Liu S, Chen R, Wang N, Liu Z, et al. Helicobacter pylori upregulates TRPC6 via Wnt/β-catenin signaling to promote gastric cancer migration and invasion. Onco Targets Ther. 2019; 12: 5269-5279.
Tsay FW, Hsu PI. H. pylori infection and extra-gastroduodenal diseases. J Biomed Sci. 2018; 25: 65.
Fischbach W, Malfertheiner P. Helicobacter Pylori Infection. Dtsch Arztebl Int. 2018; 115: 429-436.
Sunada S, Saito H, Zhang D, Xu Z, Miki Y. CDK1 inhibitor controls G2/M phase transition and reverses DNA damage sensitivity. Biochem Biophys Res Commun. 2021; 550: 56-61.
Huang Z, Zhang S, Du J, Zhang X, Zhang W, Huang Z, et al. Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803. Med Sci Monit. 2020; 26: e923664.
Caruso R, Pallone F, Monteleone G. Emerging role of IL-23/IL-17 axis in H pylori-associated pathology. World J. Gastroenterol. 2007; 13: 5547-51.
Caruso R, Fina D, Paoluzi OA, Del Vecchio Blanco G, Stolfi C, Rizzo A, et al. IL-23-mediated regulation of IL-17 production in Helicobacter pylori-infected gastric mucosa. Eur J Immunol. 2008; 38: 470-478.
Liu JF, Guo D, Kang EM, Wang YS, Gao XZ, Cong HY, et al. Acute and chronic infection of H. pylori caused the difference in apoptosis of gastric epithelial cells. Microb Pathog. 2021; 150: 104717.
Nie BX, Zhao G, Yuan XF, et al. Inhibition of CDK1 attenuates neuronal apoptosis and autophagy and confers neuroprotection after chronic spinal cord injury in vivo. J Chem Neuroanat. 2022; 119: 102053.
Liu K, Zheng M, Lu R, Du J, Zhao Q, Li Z, et al. The role of CDC25C in cell cycle regulation and clinical cancer therapy: a systematic review. Cancer Cell Int. 2020; 20: 213.
Yan J, Hao C, DeLucia M, Swanson S, Florens L, Washburn MP, et al. CyclinA2-Cyclin-dependent Kinase Regulates SAMHD1 Protein Phosphohydrolase Domain. J Biol Chem. 2015; 290: 13279-92.
Dai Y, Jin F, Wu W, Kumar SK. Cell cycle regulation and hematologic malignancies. Blood Sci. 2019; 1: 34-43.

No competing interests reported.

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H. pylori infection promotes the proliferation of gastric cancer via CDK1 expression

Status:

Version 1

Abstract

Figures

Introduction

Materials and methods

Tissue samples

Cell culture

Cell transfection

QRT-PCR assay

Assays of migration and invasion

Western blot analysis

Inflammatory factors analysis

Apoptosis analysis

Immunohistochemistry analysis

Statistical analysis

Results

Analysis of CDK1 in GC tissues

H. pylori infection influences CDK1 expression

H. pylori regulates CDK1 expression confirmed by siRNA knockdown

The inflammation induced by H. pylori through CDK1

The apoptosis of GC cell regulated by H. pylori through CDK1

Analysis of cell cycle proteins

Discussion

Declarations

References

Additional Declarations

Status:

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