摘要
目的探究miRNA-135a靶向调控HOXA10蛋白对于乳腺癌细胞生物学特性的影响。方法收集58例乳腺癌患者癌组织及癌旁正常组织,qRT-PCR检测miRNA-135a以及HOXA10在乳腺癌组织及相应的癌旁组织的表达情况;采用MCF-7乳腺癌细胞作为乳腺癌模型细胞,通过脂质体转染法转染乳腺癌细胞系MCF-7,从而构建miRNA-135a过表达细胞模型及空载体组MCF-7-NC,并以MCF-7细胞作为空白对照组;对于转染前后细胞的生物学特性进行研究,主要包括采用CCK-8法以及平板克隆的方法对转染前后细胞的增殖情况进行检测;采用划痕实验以及Transwell侵袭实验分别对于转染前后细胞的侵袭和迁移能力进行检测;使用生物信息学网站StarBase预测miRNA-135a可靶向调控的蛋白。采用qRT-PCR法检测转染前后HOXA10的mRNA表达水平;Western blot法检测细胞转染前后HOXA10蛋白的表达。结果qRT-PCR结果显示,乳腺癌组织中miRNA-135a的表达水平高于正常组织,且与患者肿瘤分化程度、浸润深度、Her2状态、肿瘤分期以及淋巴结转移密切相关,乳腺癌组织HOXA10表达水平低于正常组织(P<0.05);相比于未转染的空白对照组,转染miRNA-135a的细胞增殖、侵袭、迁移能力相比于空白对照组显著增强(P<0.05),空载体组相比于空白对照组差异无统计学意义。StarBase网站预测结果显示,miRNA-135a可与HOXA10互补结合;此外,与空白对照组相比,转染miRNA-135a的MCF-7乳腺癌细胞HOXA10的mRNA以及蛋白的表达水平明显降低(P<0.05),空载体组的表达水平无显著变化。结论miRNA-135a在乳腺癌组织中高表达,可能通过靶向抑制HOXA10蛋白的表达,从而促进乳腺癌细胞的侵袭以及迁移能力。
Objective To investigate the effect of miRNA-135a targeted regulation of HOXA10 protein on the biological characteristics of breast cancer cells.Methods MCF-7 breast cancer cells were adopted as breast cancer model cells and transfected with breast cancer cell line MCF-7 by liposome transfection to construct the mirna-135a overexpressed cell model.The biological characteristics of cells before and after transfection were observed,including the detection of cell proliferation before and after transfection by CCK-8 method.The migration and invasion abilities of the cells before and after transfection were detected by scratch test and transwell invasion test.The expression levels of HOXA10 mRNA before and after transfection were detected by qRT-PCR,moreover,the expression levels of HOXA10 protein before and after transfection were detected by Western Blot.Results The results of qRT-PCR showed that the expression levels of mirna-135a in breast cancer tissues were higher than those in normal tissues,which were closely related to the degree of tumor differentiation,depth of invasion,Her2 status,tumor stage and lymph node metastasis,moreover,and the expression levels of HOXA10 in breast cancer tissues were significantly lower than those in normal tissues(P<0.05).As compared with those in blank control group without transfection,the proliferation,invasion and migration ability in transfected mirna-135a cells were significantly increased(P<0.05),however,there were no significant differences between the empty vector group and blank control group.StarBase predicted that mirna-135a could bind with HOXA10 in a complementary way.In addition,as compared with those in blank control group,the expression levels of HOXA10 mRNA and protein in McF-7 breast cancer cells transfected with mirna-135a were significantly decreased(P<0.05),while the expression levels in empty vector group were not significantly changed.Conclusion The expression levels of miRNAa-135a are increased in breast cancer tissues,which may promote the invasion and migration of breast cancer cells by targetedly nhibiting the expression of HOXA10 protein.
作者
潘虹
余小霞
普艳红
袁瑞仙
PAN Hong;YU Xiaoxia;PU Yanhong(Department of Pathology,Qujing Hospital for Maternal and Child Health,Yunnan,Qujing 655000,China;不详)
出处
《河北医药》
CAS
2021年第6期810-814,共5页
Hebei Medical Journal
