PIK3CD-AS1对肾癌细胞增殖和凋亡的影响

Effect of PIK3CD-AS1 on proliferation and apoptosis of renal cancer cells

目的:探讨长链非编码RNA PIK3CD-AS1在肾癌细胞与正常肾脏细胞中的差异表达及其对肾癌细胞增殖和凋亡能力的影响,并寻找与PIK3CD-AS1功能相关的基因。方法:运用qRT-PCR检测PIK3CD-AS1在正常肾脏细胞(HK-2)与肾癌细胞(769-P、786-O、A498)中的表达差异;将PIK3CD-AS1过表达载体转染入肾癌769-P细胞,构建PIK3CD-AS1稳定过表达的细胞株,通过CCK-8实验及流式细胞术检测PIK3CD-AS1过表达对肾癌细胞增殖和凋亡能力的影响,通过凋亡相关基因表达谱芯片寻找与PIK3CD-AS1功能可能相关的基因。结果:与正常肾脏细胞HK-2相比,PIK3CD-AS1在肾癌细胞(769-P、786-O、A498)中表达明显下降(P<0.01或P<0.05)。成功构建PIK3CD-AS1过表达细胞株,经qRT-PCR验证,实验组细胞中PIK3CD-AS1表达量较阴性对照明显升高(P<0.05)。增殖实验及凋亡实验表明,实验组细胞的增殖能力显著下降(P<0.01),同时细胞凋亡显著增多(P<0.01)。凋亡基因表达谱芯片筛选结果显示,与PIK3CD-AS1相互作用进而促进肾癌细胞凋亡的基因有DFFA、BCL2L1、CASP9。结论:PIK3CD-AS1可能为抑癌基因,在肾癌细胞中呈现低表达状态。PIK3CD-AS1可抑制肾癌细胞的增殖并促进凋亡,其促凋亡功能可能与CASP9、DFFA、BCL2L1基因有关。

Objective： To investigate the expression of lncRNA PIK3 CD-AS1 in renal cancer cells and its effect on the proliferation and apoptosis of renal cancer cells,and the genes related to the function of PIK3 CD-AS1 were explored. Methods： The expression of PIK3 CD-AS1 in normal kidney cells（ HK-2）and renal cancer cells（ 769-P,786-O,A498） was detected by real-time fluorescence quantitative PCR（ qRT-PCR）. The PIK3 CD-AS1 overexpression vector was transfected into renal cancer cells（ 769-p） to construct a PIK3 CD-AS1 stable overexpression cell line. The effect of PIK3 CD-AS1 on the proliferation and apoptosis of renal cancer cells was detected by CCK-8 assay and flow cytometry. The apoptosis-related gene expression profile chip was used to screen genes that may be related to PIK3 CD-AS1 function.Results： The expression of PIK3 CD-AS1 was significantly decreased in kidney cancer cells（ 769-P,786-O,A498） compared with normal renal cell HK-2（ P〈0. 01 or P〈0. 05）. The PIK3 CD-AS1 overexpression cell line was successfully constructed and verified by qRT-PCR. The expression of PIK3 CD-AS1 in the experimental group was significantly higher than that of the negative control（ P〈0. 05）. The proliferation and apoptosis experiments showed that the proliferation ability of the cells in the experimental group was significantly decreased（ P〈0. 01）,and the apoptosis was significantly increased（ P〈0. 01）. The results of gene expression microarray screening showed that the genes interacting with PIK3 CD-AS1 and promoting apoptosis are DFFA,BCL2 L1,and CASP9. Conclusion： PIK3 CD-AS1 may be a tumor suppressor gene and is lowly expressed in renal cancer cells. PIK3 CD-AS1 can inhibit the proliferation of renal cancer cells and promote apoptosis,and its pro-apoptotic function may be related to CASP9,DFFA,BCL2 L1 genes.