沉默长链非编码RNA HOTAIR对膀胱癌细胞增殖、凋亡的影响及其机制

Effect of silencing LncRNA HOTAIR on proliferation and apoptosis of bladder cancer cell line 253J and its mechanism

目的观察沉默长链非编码RNA(LncRNA) HOTAIR对膀胱癌细胞增殖、凋亡的影响,并探讨其作用机制。方法取适量对数生长期膀胱癌253J细胞,随机分为si-HOTAIR干扰组、空载体组、对照组,si-HOTAIR干扰组、空载体组分别转染si-HOTAIR(可沉默HOTAIR表达)及空载体阴性序列,对照组不做任何处理。转染4 h,采用CCK-8法检测各组细胞增殖情况,采用Hoechst法检测各组细胞凋亡情况,采用实时荧光定量PCR法检测各组细胞Notch信号通路蛋白1(Notch1)、上皮细胞黏附分子(Ep CAM)、趋化因子受体2(CXCR2)、上皮性黏附蛋白(Ecadherin)、波形蛋白(Vimentin)、基质金属蛋白酶2(MMP-2)和α平滑肌肌动蛋白(α-SMA) mRNA表达。结果转染4 h,si-HOTAIR干扰组、空载体组、对照组细胞增殖能力分别为0. 38±0. 02、0. 89±0. 01、0. 89±0. 02,与对照组、空载体组比较,si-HOTAIR组细胞增殖能力明显下降(P均<0. 05); si-HOTAIR干扰组、空载体组、对照组凋亡细胞数分别为(33. 56±2. 26)、(13. 69±1. 25)、(12. 85±1. 35)个,与对照组、空载体组比较,si-HOTAIR组凋亡细胞数升高(P均<0. 05);与对照组、空载体组比较,si-HOTAIR组Notch1、Ep CAM、CXCR2、Vimentin、MMP-2和α-SMA mRNA相对表达量均明显下降,E-cadherin mRNA相对表达量明显升高(P均<0. 05)。结论沉默LncRNA HOTAIR表达可抑制膀胱癌细胞增殖并促进其凋亡,其作用机制可能与抑制Notch1、Ep CAM、CXCR2、Vimentin、MMP-2和α-SMA表达,促进E-cadherin表达有关。

Objective To observe the effects of silencing long-chain non-coding RNA(LncRNA)HOTAIR on the proliferation and apoptosis of bladder cancer cell line 253J and to explore its mechanism.Methods 253J cells in logarithmic phase were divided into the si HOTAIR interference group,empty vector group,and control group.The cells in the si HOTAIR interference group and empty vector group were transfected with siHOTAIR(which could silence the expression of HOTAIR)and the negative sequence of the empty vector,respectively.No treatment was done in the control group.CCK8 assay was used to observe the proliferation of cells in each group after 4-hour transfection,and Hoechst method was used to observe the apoptosis in each group.The mRNA expression of Notch1,EpCAM,CXCR2,E-cadherin,vimentin,matrix metalloproteinase 2(MMP-2)andα-SMA in each group was detected by real-time fluorescence quantitative PCR.Results At 4 h after transfection,the OD values of the si HOTAIR interference group,empty vector group and control group were 0.38±0.02,0.89±0.01,and 0.89±0.02,respectively.Compared with the empty vector group and control group,the OD value of the si HOTAIR group decreased(P<0.05).At 4 h after transfection,the number of apoptotic cells in the si HOTAIR interference group,empty vector group and control group were 33.56±2.26,13.69±1.25,and 12.85±1.35,respectively.The number of apoptotic cells in the si HOTAIR group was higher than that of the control group and empty vector group(P<0.05).Compared with the control group and empty vector group,the relative mRNA expression of Notch1,EpCAM,CXCR2,Vimentin,MMP-2,andα-SMA decreased,but E-cadherin mRNA expression increased in the si HOTAIR interference group(all P<0.05).Conclusion Silencing HOTAIR expression can inhibit the proliferation and promote the apoptosis of bladder cancer cells by inhibiting the expression of Notch1,EpCAM,CXCR2,Vimentin,MMP-2,andα-SMA and promoting the expression of E-cadherin.