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[小干扰RNA介导的BIRC6沉默对肾癌786-O细胞凋亡和自噬的影响]

[Effect of small interfering RNA-mediated BIRC6 silencing on apoptosis and autophagy of renal cancer 786-O cells].

作者信息

Zhong Kaihua, Chen Dong, Wu Zhiming, Wang Xiaohong, Pan Bin, Chen Nanhui, Zhong Weifeng

机构信息

Department of Urology, Meizhou People's Hospital, Meizhou 514031, China.

Department of Urology, Sun Yat-sen Cancer Center, Guangzhou 510060, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2020 Nov 30;40(11):1651-1655. doi: 10.12122/j.issn.1673-4254.2020.11.18.

Abstract

OBJECTIVE

To study the expression of BIRC6 in renal cancer tissues and investigate the effect of BIRC6 silencing on apoptosis and autophagy of 786-O cells.

METHODS

Twenty surgical specimens of renal cancer tissues and adjacent renal tissues were collected from Meizhou People's Hospital between February, 2016 and December, 2018 for detection of BIRC6 protein expression using immunohistochemistry. Renal cancer 786-O cells were transfected with a control small interfering RNA (siRNA) or BIRC6 siRNA lipofectamine 2000, and the changes in cell proliferation and apoptosis following 5-FU treatment were assessed using CCK8 assay and flow cytometry; the expressions of autophagy-related proteins Beclin and LC3A/B were detected by Western blotting.

RESULTS

The expression of BIRC6 protein was significantly higher in renal cancer tissues than in the adjacent renal tissues. Western blotting showed that siRNA-mediated silencing of BIRC6 significantly lowered the expression of BIRC6 in 786-O cells. In the cells with BIRC6 silencing, treatment with 12.5, 25, 50, 100 and 200 μg/mL 5-FU resulted in significantly higher proliferation inhibition rates than in the cells transfected with the control siRNA ( < 0.01). BIRC6 silencing also significantly increased the apoptosis rate of 786-O cells following 5-FU treatment ( < 0.01). The results of Western blotting showed that BIRC6 silencing significantly lowered the protein expressions of Beclin and LC3A/B in 786-O cells.

CONCLUSIONS

Interference of BIRC6 mediated by siRNA can inhibit autophagy and promote 5-FU-induced apoptosis to enhance the sensitivity of 786-O cells to 5-FU.

摘要

目的

研究BIRC6在肾癌组织中的表达情况,并探讨BIRC6沉默对786-O细胞凋亡和自噬的影响。

方法

收集2016年2月至2018年12月期间梅州人民医院的20例肾癌组织手术标本及癌旁肾组织,采用免疫组织化学法检测BIRC6蛋白表达。将肾癌786-O细胞用对照小干扰RNA(siRNA)或BIRC6 siRNA脂质体2000转染,采用CCK8法和流式细胞术评估5-FU处理后细胞增殖和凋亡的变化;通过蛋白质免疫印迹法检测自噬相关蛋白Beclin和LC3A/B的表达。

结果

肾癌组织中BIRC6蛋白的表达明显高于癌旁肾组织。蛋白质免疫印迹法显示,siRNA介导的BIRC6沉默显著降低了786-O细胞中BIRC6的表达。在BIRC6沉默的细胞中,用12.5、25、50、100和200μg/mL 5-FU处理后,增殖抑制率明显高于转染对照siRNA的细胞(<0.01)。BIRC6沉默还显著提高了5-FU处理后786-O细胞的凋亡率(<0.01)。蛋白质免疫印迹法结果显示,BIRC6沉默显著降低了786-O细胞中Beclin和LC3A/B的蛋白表达。

结论

siRNA介导的BIRC6干扰可抑制自噬并促进5-FU诱导的凋亡,从而增强786-O细胞对5-FU的敏感性。

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