慢病毒RNA干扰GRP78对肾癌细胞生物学行为的影响研究

付伟金; 吕佰豪; 顾恺龙

doi:10.13201/j.issn.1001-1420.2016.12.018

慢病毒RNA干扰GRP78对肾癌细胞生物学行为的影响研究

- 广西医科大学第一附属医院泌尿外科(南宁, 530021)
基金项目:
广西自然科学基金资助项目(编号2012GXNSFBA053087)

详细信息

通讯作者: 付伟金,E-mail:fuwj66@aliyun.com

中图分类号: R737.11

收稿日期: 2016-04-22

Research on the effect of lentivirus-siRNA-GRP78 on biological behavior of renal cell carcinoma

- Department of Urology, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China

More Information

Corresponding author: FU Weijin, E-mail: fuwj66@aliyun.com

Received Date: 22 April 2016

摘要

摘要: 目的:探讨慢病毒RNA干扰GRP78蛋白后对肾癌细胞生物学行为的影响。方法:构建慢病毒靶向siRNA-GRP78干扰载体进入肾癌细胞株。转染48 h后使用Realtime-PCR和Western bloting 检测GRP78基因和蛋白表达,MTT法和transwell法分别检测肾癌细胞增殖和侵袭力。结果:转染48 h后,Realtime-PCR和Western bloting检测证实siRNA1抑制GRP78基因和蛋白效率最佳。SiRNA1-GRP78可抑制肾癌细胞增殖和降低细胞侵袭力,促进细胞凋亡,阻滞细胞于G1期。结论:慢病毒RNA干扰可有效抑制肾癌细胞中GRP78基因和蛋白表达,GRP78表达下调影响肾癌细胞增殖和降低细胞侵袭力,诱导细胞凋亡,阻滞细胞于G1周期。GRP78可能成为肾癌治疗新靶点。
- 慢病毒RNA干扰 /
- GRP78 /
- 增殖 /
- 侵袭力 /
- 细胞凋亡 /
- 细胞周期
Abstract: Objective: To investigate the effect of lentivirus-siRNA-GRP78 on biological behavior of renal cell carcinoma (RCC).Method: According to GRP78 gene sequences, we constructed three siRNA -GRP78 sequences and one scrambled siRNA sequence, which were transfected into RCC cell line 786-0. After transferring for 48h, we identified the expression level of GRP78 mRNA and protein by realtime-PCR and Western blot for selecting the optimal inhibiting siRNA-sequence. MTT, transwell, Annexin V, PI were used respectively to detect the ability of cell proliferation, cell invasion, cell apoptosis and cell cycle.Result: Subsequent experiments have focused on the siRNA1 because it was the most effective in inhibiting GRP78 mRNA and protein expression. SiRNA1-GRP78 may inhibit the cell proliferation, attenuate cell invasion, induce cell apoptosis and block cell cycle in G1 stage.Conclusion: Lentivirus-siRNA-GRP78 can effectively inhibit the expression GRP78 mRNA and protein, and the down-regulation of GRP78 expression could inhibit cell proliferation, attenuate cell invasion, induce cell apoptosis, block cell cycle in G1 stage. GRP78 may be a promising therapeutic target for RCC.
- lentivirus-siRNA /
- GRP78 /
- proliferation /
- invasion /
- cell apoptosis /
- cell cycle

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