| 张靖华,吴秋杰,刘金瑞.HAI-1对肾细胞癌细胞自噬及凋亡调控的初步研究[J].中国肿瘤,2020,29(9):715-720. |
| HAI-1对肾细胞癌细胞自噬及凋亡调控的初步研究 |
| Effect of HAI-1 on Autophagy and Apoptosis in Renal Cell Cancer Cells |
| 中文关键词 修订日期：2020-03-05 |
| DOI：10.11735/j.issn.1004-0242.2020.09.A013 |
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| 中文关键词: 肾细胞癌 肝细胞生长因子激活因子1型抑制因子 自噬 凋亡 |
| 英文关键词:renal cell carcinoma hepatocyte growth factor activator type 1 inhibitor autophagy apoptosis |
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| 中文摘要: |
| 摘 要：［目的］ 研究肝细胞生长因子激活因子1型抑制因子（hepatocyte growth factor activator inhibitor type 1，HAI-1）对肾细胞癌细胞自噬和凋亡的影响，探讨其发挥作用的可能机制。［方法］ QRT-PCR检测HAI-1在肾细胞癌组织和细胞系中的表达水平，选择低表达的肾细胞癌细胞系进行HAI-1过表达质粒的转染，分为对照组（NC组）和过表达质粒组（OE-HAI-1组）。QRT-PCR检测NC组和OE-HAI-1组细胞中HAI-1的表达水平；MTS实验检测NC组和OE-HAI-1组细胞的增殖率；Western blot法检测HAI-1对肾细胞癌细胞中自噬相关蛋白表达水平的影响；流式细胞仪检测NC组和OE-HAI-1组细胞凋亡率；Western blot法检测NC组和OE-HAI-1组细胞中caspase3、Bax和Bcl-2蛋白的表达水平。［结果］ QRT-PCR结果显示HAI-1在肾细胞癌组织中的表达低于其在癌旁组织中的表达，HAI-1在肾细胞癌细胞系ACHN、786-O和A-498中的表达低于其在肾小管上皮细胞HK-2中的表达（P＜0.05）。选择HAI-1表达最低的肾细胞癌细胞系786-O转染HAI-1过表达质粒，qRT-PCR结果显示OE-HAI-1组细胞中HAI-1的表达增加（P＜0.05）；与NC组相比，OE-HAI-1组786-O细胞增殖能力下降。与NC组比较，OE-HAI-1组和加入自噬激活剂Rapamycin的OE-HAI-1+Rapamycin组细胞中Beclin1和LC3-Ⅱ蛋白表达降低，LC3-Ⅰ蛋白表达增加。与OE-HAI-1组相比，OE-HAI-1+Rapamycin组Beclin1和LC3-Ⅱ蛋白表达增加，LC3-Ⅰ蛋白表达降低。与NC组相比，OE-HAI组细胞凋亡增加，细胞中caspase3和Bax蛋白表达增加，Bcl-2蛋白的表达降低。［结论］ HAI-1在肾细胞癌组织和细胞系中表达降低，并促进肾细胞癌细胞自噬和凋亡的发生。HAI-1可能是治疗肾细胞癌的候选分子靶标。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the effect of hepatocyte growth factor activator inhibitor type 1(HAI-1) on autophagy and apoptosis in renal cell carcinoma cells，and its possible mechanisms. ［Methods］ The expression levels of HAI-1 were detected with qRT-PCR in renal cell carcinoma tissues and cell lines. The HAI-1 plasmids(OE-HAI-1 group) or blank plasmids(NC group) were transfected in low-expressing renal cell carcinoma cells. MTS was used to detect the cell proliferation rate；Western blot method was used to detect the effect of HAI-1 on autophagy-related proteins expression in renal cell carcinoma cells；flow cytometry was used to detect apoptosis rates in NC group and OE-HAI-1 group. The expressions of caspase3，Bax and Bcl-2 proteins were detected with Western blot. ［Results］ The qRT-PCR results showed that the expression of HAI-1 in renal cell carcinoma tissues was lower than that in cancer adjacent tissues，and the expression of HAI-1 in renal cell carcinoma cell lines ACHN，786-O，and A-498 was lower than that in renal tubular epithelial HK-2 cells(P<0.05). The renal cell carcinoma cell line 786-O with the lowest HAI-1 expression was selected to transfect the HAI-1 overexpression plasmid. The qRT-PCR results showed that HAI-1 expression was increased in the OE-HAI-1 cells(P<0.05). Compared with the NC group，the expressions of Beclin1 and LC3-Ⅱ proteins were decreased，and the expression of LC3-Ⅰ protein was increased in the OE-HAI-1 cells and OE-HAI-1 + Rapamycin group. Compared with the OE-HAI-1 group，the expressions of Beclin1 and LC3-Ⅱ proteins were increased and the expression of LC3-Ⅰ protein was decreased in the OE-HAI-1 + Rapamycin group. Compared with the NC group，the apoptosis of the OE-HAI group was increased，the expression of caspase3 and Bax protein were increased，and the expression of Bcl-2 protein was decreased. ［Conclusion］ HAI-1 expression is downregulated in renal cell carcinoma tissues and cell lines，resulting in the autophagy and apoptosis in renal cell carcinoma cells. HAI-1 may be a candidate molecular target for the treatment of renal cell carcinoma. |
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