HOXB5基因低表达对膀胱癌细胞增殖、凋亡、侵袭的影响及机制

Effects of low expression of HOXB5 gene on proliferation,apoptosis,and invasion of bladder cancer cells

目的探讨HOXB5基因低表达对膀胱癌细胞增殖、凋亡、侵袭的影响及机制。方法常规培养人膀胱癌细胞(T24、5637、BIU-87细胞)及正常膀胱上皮细胞(SV-HUC-1细胞),用RT-PCR检测各细胞中HOXB5 mRNA表达,T24细胞中HOXB5相对表达量表达最高,将T24细胞作为实验细胞。将T24细胞分为空白组、NC组与转染组,空白组常规培养,将si-NC质粒、si-HOXB5质粒转染NC组、转染组,用RT-PCR检测转染效率。MTT法测算细胞增殖抑制率,用流式细胞术测算细胞凋亡率,用Transwell小室实验检测细胞侵袭能力,用Western blotting法检测细胞内人类eag相关基因(HERG1)、转化生长因子β(1 TGF-β_(1))、血管内皮生长因子(VEGF)蛋白表达。结果与空白组、NC组比较,转染组细胞增殖抑制率、凋亡率高,穿膜细胞数少,HERG1、TGF-β_(1)、VEGF蛋白相对表达量低(P均<0.05);空白组与NC组以上指标比较差异无统计学意义(P均>0.05)。结论HOXB5基因低表达可抑制膀胱癌细胞增殖、侵袭,促进其凋亡;其作用机制可能与降低HERG1、TGF-β_(1)、VEGF蛋白表达有关。

Objective To explore the effects of HOXB5 gene low expression on the proliferation,apoptosis,and in‑vasion of bladder cancer cells and the mechanism.Methods We routinely cultured human bladder cancer cells(T24,5637,BIU-87 cells)and normal bladder epithelial cells(SV-HUC-1 cells),and used RT-PCR to detect the expression of HOXB5 mRNA in the cells.The relative expression level of HOXB5 was the highest in T24 cells,and then T24 cells were used as experimental cells.We divided T24 cells into the blank group,NC group,and transfection group.The cells in the blank group were routinely cultured.The si-NC plasmid and si-HOXB5 plasmid were transfected into the NC group and the transfection group.The transfection efficiency was calculated by RT-PCR.MTT was used to measure the inhibition rate of cell proliferation,flow cytometry was applied to measure the rate of apoptosis,Transwell chamber experiment was used to detect cell invasion ability,and Western blotting was used to detect the expression of human eag-related genes(HERG1),transforming growth factor-β_(1)(TGF-β_(1)),and vascular endothelial growth factor(VEGF)protein in the cells.Results Compared with the blank group and NC group,the transfection group had higher cell proliferation inhibition rate,higher apoptosis rate,fewer penetrating cells,and lower relative expression levels of HERG1,TGF-β_(1),and VEGF protein(all P<0.05).There was no significant difference in the above indicators between the blank group and the NC group(all P>0.05).Conclusion The low expression of HOXB5 gene can inhibit the proliferation and invasion of bladder cancer cells,and promote their apoptosis,and its mechanism of action may be related to the reduction of HERG1,TGF-β_(1),and VEGF protein expression.