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Received:April 05, 2022 Published Online:January 20, 2023
Received:April 05, 2022 Published Online:January 20, 2023
中文摘要: 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+10pm/mlAPJ拮抗剂Apelin13(F13A)]、联合组(MCF-7细胞+10pm/mlF13A+HER-3敲低)。采用qRT-PCR、CCK-8、流式细胞仪检测各组细胞HER-3mRNA的表达、细胞活力及凋亡率;给予0、2、4、6、8Gy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-3mRNA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96h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、8Gy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。
中文关键词: 乳腺癌 人表皮生长因子受体-3 放射抵抗 G蛋白偶联受体
Abstract: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 +10pm/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-1α were 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-1α in 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 breast cancer cells, accelerate apoptosis, and increase the sensitivity of breast cancer radiotherapy to a certain extent. The mechanism may be related to the inhibition of the activity of Apelin/APJ signaling pathway.
keywords: Breast cancer Human epidermal growth factor receptor-3 Radioresistance G protein coupled receptor
文章编号: 中图分类号:R737.9 文献标志码:A
基金项目:国家自然科学基金资助项目(81762062)
Author Name | Affiliation |
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 |
Author Name | Affiliation |
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 |
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