长链非编码RNA HOXC13-AS在肝细胞癌中的表达及对细胞增殖、侵袭的影响

Expression of long non-coding RNA HOXC13-AS in hepatocellular carcinoma and its effects on cell proliferation and invasion

目的观察长链非编码RNA HOXC13反义RNA(RNA HOXC13-AS)在肝细胞癌中的表达,探讨其对肝癌细胞增殖和侵袭的影响。方法实时荧光定量PCR(qRT-PCR)检测肝癌细胞株及正常肝细胞株中HOXC13-AS相对表达水平。体外培养HepG2细胞,设置空白对照组(不转染)、LV-NC组(转染携带HOXC13-AS-NC慢病毒)、LV-HOXC13-AS-shRNA组(转染携带HOXC13-AS-shRNA慢病毒),CCK-8法检测各组细胞增殖情况,Transwell法及细胞划痕实验检测各组肝癌细胞侵袭及迁移能力,蛋白免疫印记(WB)法检测增殖与侵袭相关蛋白表达水平。结果与人正常肝细胞株L02(0.22±0.03)比较,肝癌细胞株HepG2(1.02±0.04)、SMMC-7721(0.71±0.03)、HUH7(0.59±0.03)、SNU-182(0.41±0.03)中HOXC13-AS的表达水平显著升高,差异均有统计学意义(均P<0.05),以肝癌细胞HepG2中HOXC13-AS相对表达水平最高。与空白对照组、LV-NC组比较,LV-HOXC13-AS-shRNA组中HepG2细胞增殖、迁移与侵袭能力及基质金属蛋白酶-2、Ki-67、细胞周期蛋白D1(Cyclin D1)相对表达水平明显降低,而组织金属蛋白酶抑制剂-2蛋白相对表达水平明显升高,差异均有统计学意义(均P<0.05)。结论HOXC13-AS在肝癌细胞中高表达,HOXC13-AS低表达可抑制HepG2细胞的增殖、侵袭。

Objective To observe the expression of long non-coding RNA HOXC13-AS in hepatocellular carcinoma and to explore its effects on the proliferation and invasion of hepatocellular carcinoma.Methods The relative expression of HOXC13-AS in hepatoma cell line and normal hepatoma cell line was detected by real-time fluorescent quantitative PCR(qRT-PCR).HepG2 cells were cultured in vitro,and divided into the blank control group(not transfected),LV-NC group(transfected with HOXC13-AS-NC lentivirus),and LV-HOXC13-AS-shRNA group(transfected with HOXC13-AS-shrna lentivirus).CCK-8 method was used to detect cell proliferation.Transwell method and cell scratch test were used to detect the invasion and migration of hepatoma cells,and western blot(WB)was used to detect the expressions of proliferation and invasion related proteins.Results Compared with human normal hepatocyte line L02 group(0.22±0.03),the expression levels of HOXC13-AS in hepatoma cell lines HepG2(1.02±0.04),SMMC-7721(0.71±0.03),HUH7(0.59±0.03)and SNU-182(0.41±0.03)increased significantly(P<0.05),and the relative expression level of HOXC13-AS in HepG2 was the highest;compared with the blank control group and LV-NC group,the proliferation,migration and invasion ability of HepG2 cells and the relative expression levels of matrix metalloproteinase-2,Ki-67 and Cyclin D1 protein in LV-HOXC13-ASshRNA group were significantly reduced,however,the relative expression level of tissue inhibitor of metalloproteinases-2 protein was significantly increased(P<0.05).Conclusion HOXC13-AS was highly expressed in hepatoma cells,and the low expression of HOXC13-AS could inhibit the proliferation and invasion of HepG2 cells.