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| Effect of Silencing RALA Gene Expression on the Proliferation, Migration and Apoptosis of Prostate Cancer Cells |
| WENG Wubin, LIU Changming, ZHANG Jiabin, LI Guomin, XUE Qingping, LIN Ningfeng, CHEN Guangbing |
| Mindong Hospital Affiliated to Fujian Medical University, Fu'an 355000, China |
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Abstract Objective: To investigate the effects of small interfering RNA (siRNA) inhibition of RALA gene expression on the proliferation, migration and regulation of prostate cancer cells and its possible mechanisms. Method: RT-PCR was used to detect the expression levels of RALA mRNA, DU145, PC-3 and LNCap cellsin prostate cancer. The RALA-siRNA was transfected into somatic cultured DU145 cell lines by LipofectamineTM 2000, and the effect of RALA-siRNA on the proliferation of DU145 cell lines was detected by MTT assay; the effect of RALA-siRNA on the migration ability of DU145 cell lines was detected by Transwell assay; the flow cytometry was used to assay the effect of RALA-siRNA on apoptosis of DU145 cell line. The expression levels of RALA gene and protein in DU145 cell line after silencing RALA gene were detected by RT-PCR and Western blot. Result: The RT-PCR results suggested that the expression levels of RALA mRNA in DU145 cell line were higher than those in PC-3 and LNCap cell lines (P<0.05), and their relative expression levels were (0.83±0.02), (0.37±0.07) and (0.41±0.01). MTT assay results showed that the expression levels of RALA mRNA after transfection with RALA- siRNA significantly inhibited the proliferation of DU145 cell line, the difference was statistically significant compared with the negative-siRNA and blank control groups (P<0.05); the results of Transwell assay showed that the number of membrane penetrating cells in the RALA-siRNA group was lower than those in the negative-siRNA and blank control groups (P<0.05). The results of flow cytometry showed that the apoptosis rate in the RALA-siRNA group was significantly higher than those in the negative-siRNA and blank control groups (P<0.05). Western blot assay results suggested that the expression of RALA protein in the RALA-siRNA group was significantly inhibited compared with those in the negative-siRNA and blank control groups (P<0.05). Conclusion: The expression level of RALA gene is higher in prostate cancer DU145 cell line, and silencing RALA gene can restrict the proliferation and migration ability of DU145 cells, suggesting that RALA may be a potential target in prostate cancer therapy.
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Received: 08 October 2022
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