This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research article
Li Li
Corresponding Author
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Background: Dyhidrofolate reductase (DHFR) is involved in the DNA synthesis and is expressed highly in platinum resistant ovarian carcinoma tissue . The relationship between DHFR gene and platinum drug resistance in ovarian carcinoma (OC) is still unclear.
Methods: To design targeting hairpin siRNA of DHFR gene, and constructed lentiviral vector carrying DHFR gene, then though flow cytometry、high performance liquid chromatography and electron microscope, DHFR gene expression was down-regulating to investigate the biological behaviour of DHFR in OC.
Results: DHFR mRNA expression in ovarian carcinoma was higher than normal ovarian tissues, in patients with omentum metastasis was lower than that in patients without omentum metastasis, in patients with CR for treatment was lower than that of patients with SD or PD or PR, cisplatin-sensitive patients was lower than that in cisplatin-multidrug resistant cases. The median survival period of patients with DHFR mRNA expression lower than 0.331(Youden index) was 16.4 months, but of that higher than 0.331 was 44.5 months. COX multivariate analysis showed that DHFR mRNA expression was a dependent prognostic factor. The apoptosis rate of DHFR-pGCSIL-SKOV3 was obvious higher than pGCSIL-SKOV3 and SKOV3 cells at 24h and 48h(P＜0.05). GO/G1 stage rate of three groups at 24h、48h and 72h were decreasing but the G2/M and S stage rates were raising. The cells in the cisplatin concentration (2.5、5.0μg/mL)at 24h and 48h,the intracellular cisplatin content of DHFR-pGCSIL-SKOV3 cell were significantly higher than pGCSIL-SKOV3 and SKOV3 cells. The microfilament increased and gathered together, mitochondrial structure also change obviously under no drug. But there were rarely microfilament in three group cells induced by IC50 cisplatin concentration(4.4、5.5、4.9µg/ml)at 24 and 48 h, while at 72 h, there are obvious increase and inordinate microfilament.
Conclusion: DHFR silencing inhibits cell growth and cisplatin resistance, there were certain contact between resistance increases with microfilament gathered and the change of the mitochondria.The results laid the foundation for us to investigate the molecular mechanisms of multidrug-resistance in tumor.
Keywords
DHFR, ovarian carcinoma, cisplatin-resistance, silencing
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research article
Li Li
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 23 Dec, 2019
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Dyhidrofolate reductase (DHFR) is involved in the DNA synthesis and is expressed highly in platinum resistant ovarian carcinoma tissue . The relationship between DHFR gene and platinum drug resistance in ovarian carcinoma (OC) is still unclear.
Methods: To design targeting hairpin siRNA of DHFR gene, and constructed lentiviral vector carrying DHFR gene, then though flow cytometry、high performance liquid chromatography and electron microscope, DHFR gene expression was down-regulating to investigate the biological behaviour of DHFR in OC.
Results: DHFR mRNA expression in ovarian carcinoma was higher than normal ovarian tissues, in patients with omentum metastasis was lower than that in patients without omentum metastasis, in patients with CR for treatment was lower than that of patients with SD or PD or PR, cisplatin-sensitive patients was lower than that in cisplatin-multidrug resistant cases. The median survival period of patients with DHFR mRNA expression lower than 0.331(Youden index) was 16.4 months, but of that higher than 0.331 was 44.5 months. COX multivariate analysis showed that DHFR mRNA expression was a dependent prognostic factor. The apoptosis rate of DHFR-pGCSIL-SKOV3 was obvious higher than pGCSIL-SKOV3 and SKOV3 cells at 24h and 48h(P＜0.05). GO/G1 stage rate of three groups at 24h、48h and 72h were decreasing but the G2/M and S stage rates were raising. The cells in the cisplatin concentration (2.5、5.0μg/mL)at 24h and 48h,the intracellular cisplatin content of DHFR-pGCSIL-SKOV3 cell were significantly higher than pGCSIL-SKOV3 and SKOV3 cells. The microfilament increased and gathered together, mitochondrial structure also change obviously under no drug. But there were rarely microfilament in three group cells induced by IC50 cisplatin concentration(4.4、5.5、4.9µg/ml)at 24 and 48 h, while at 72 h, there are obvious increase and inordinate microfilament.
Conclusion: DHFR silencing inhibits cell growth and cisplatin resistance, there were certain contact between resistance increases with microfilament gathered and the change of the mitochondria.The results laid the foundation for us to investigate the molecular mechanisms of multidrug-resistance in tumor.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
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