HER-3基因表达对乳腺癌细胞放射抵抗及Apelin/APJ信号通路的作用机制  

HER-3 gene expression on radioresistance and Apelin/APJ signaling pathway of breast cancer cells

作　　者：李馨如 罗成 荀欣 贾舒婷 LI Xin-ru;LUO Cheng;XUN Xin;JIA Shu-ting(Department of Oncology,The 920th Hospital of PLA Joint Logistics Support Force,Kunming,Yunnan 650000,China)

机构地区：[1]解放军联勤保障部队第九二〇医院肿瘤科,云南昆明650000

出　　处：《中国临床研究》2023年第1期12-17,共6页Chinese Journal of Clinical Research

基　　金：国家自然科学基金资助项目(81762062)。

摘　　要：目的探讨人表皮生长因子受体(HER)-3基因表达在乳腺癌细胞放射抵抗及Apelin/G蛋白偶联受体(APJ)信号通路中的作用。方法将乳腺癌细胞株处理后分为乳腺癌组(生长状态良好的MCF-7细胞)、空白组(MCF-7细胞+阴性对照慢病毒)、HER-3组(MCF-7细胞+HER-3敲低)、F13A组[MCF-7细胞+10 pm/ml APJ拮抗剂Apelin13(F13A)]、联合组(MCF-7细胞+10 pm/ml F13A+HER-3敲低)。采用qRT-PCR、CCK-8、流式细胞仪检测各组细胞HER-3 mRNA的表达、细胞活力及凋亡率;给予0、2、4、6、8 Gy放射剂量照射观察各组细胞放射敏感性;各组细胞Apelin13、APJ、血管内皮生长因子(VEGF)及低氧诱导因子(HIF)-1α蛋白的表达采用免疫印迹法检测,其与HER-3的相关性采用Pearson法分析。结果乳腺癌组、空白组、HER-3组、F13A组及联合组的HER-3 mRNA相对表达量分别为1.00±0.00、0.96±0.03、0.58±0.05、0.62±0.06及0.23±0.04(F=346.100,P<0.01);与乳腺癌组及空白组相比,HER-3组、F13A组及联合组24、48、72及96 h时MCF-7细胞活力均降低(P<0.05);各组MCF-7细胞凋亡率差异有统计学意义(F=125.200,P<0.05),联合组细胞凋亡率最高(P<0.05);照射剂量为2、4、6、8 Gy时HER-3组、F13A组及联合组细胞活性均低于乳腺癌组及空白组(P<0.05);与乳腺癌组及空白组比较,HER-3组、F13A组及联合组Apelin13、APJ、VEGF及HIF-1α蛋白表达均降低(P<0.05);HER-3分别与Apelin13、APJ、VEGF及HIF-1α呈现正相关性(P<0.05)。结论敲低HER-3基因可以抑制乳腺癌细胞活性,加快凋亡,并可一定程度上增加乳腺癌放疗敏感性,其机制可能与抑制Apelin/APJ信号通路活性相关。Objective To investigate the mechanism of human epidermal growth factor receptor-3(HER-3)gene expression on radioresistance and Apelin(APJ-endogenous ligand apelin)/APJ(G protein-coupled receptor)signaling pathway in breast cancer cells.Methods Breast cancer cells were divided into breast cancer group(MCF-7 cells in good state),blank group(MCF-7 cells+negative control lentivirus),HER-3 group(MCF-7 cells+knockdown of HER-3),F13A group[MCF-7 cells+10 pm/ml F13A(APJ-specific antagonist)]and combined group(MCF-7 cells+10 pm/ml F13A+knockdown of HER-3).The expressions of HER-3 mRNA,cell viability and apoptosis rate were respectively detected by qRT-PCR,CCK-8 and flow cytometry.Radiosensitivity was observed by 0,2,4,6 and 8 Gy radiation doses in each group.The protein expressions of Apelin13,APJ,VEGF and HIF-1αwere detected by western blot,and Pearson method was used to analyze the correlation between them and HER-3.Results The mRNA expressions of HER-3 in breast cancer group,blank group,HER-3 group,F13A group and combined group were 1.00±0.00,0.96±0.03,0.58±0.05,0.62±0.06 and 0.23±0.04,respectively(F=346.100,P<0.01).Compared with those in breast cancer group and blank group,the activity of MCF-7 cell line in HER-3 group,F13A group and combined group statistically decreased at 24-,48-,72-and 96-h(P<0.05).There was a significant difference in apoptosis rate of MCF-7 cell lines among five groups(F=125.200,P<0.05),and the apoptosis rate was the highest in combined group than those of other groups(P<0.05).When the irradiation dose was 2-,4-,6-and 8-Gy,the cell activity in HER-3 group,F13A group and combined group were significantly lower than that in breast cancer group and blank group(P<0.05).Compared with breast cancer group and blank group,the protein expressions of Apeli13,APJ,VEGF and HIF-1αin HER-3 group,F13A group and combined group decreased significantly(P<0.05).HER-3 was positively correlated with Apeli13,APJ,VEGF and HIF-1α(P<0.05),respectively.Conclusion Knockdown HER-3 gene can inhibit the activity of

关 键 词：乳腺癌 人表皮生长因子受体-3 放射抵抗 G蛋白偶联受体 

分 类 号：R737.9[医药卫生—肿瘤]