miR-96-5p靶向HOXA9促进乳腺癌细胞增殖侵袭的机制研究  

Study on the mechanism of miR-96-5p targeting HOXA9 to promote the proliferation and invasion of breast cancer cells

作　　者：吴丽华[1] 钟慕仪 黄珂铭[1] 王西跃 罗锐娟 陈丽娟 林正权[1] 袁惠玲 WU Lihua;ZHONG Muyi;HUANG Keming;WANG Xiyue;LUO Ruijuan;CHEN Lijuan;LIN Zhengquan;YUAN Huiling(Department of Mammary Gland,the 10th Affiliated Hospital of Southern Medical University,Dongguan People’s Hospital,Dongguan,Guangdong 523000,China)

机构地区：[1]南方医科大学第十附属医院/东莞市人民医院乳腺科,广东东莞523000

出　　处：《中国优生与遗传杂志》2024年第2期228-236,共9页Chinese Journal of Birth Health & Heredity

基　　金：2022年东莞市社会发展科技重点项目(20221800906302)。

摘　　要：目的探究miR-96-5p靶向同源盒基因A9(HOXA9)促进乳腺癌细胞增殖侵袭的机制。方法收集2022年9—12月经手术切除的60例乳腺癌、癌旁组织样本及患者完整临床资料,miRNA微阵列分析2对乳腺癌与癌旁组织中差异表达的miRNA。qRT-PCR检测乳腺癌、癌旁组织及乳腺癌细胞系miR-96-5p表达水平,分析miR-96-5p表达与临床病理特征的关系。通过MTT、克隆形成、EdU法、划痕、Transwell实验与体内异种移植实验检测miR-96-5p对乳腺癌细胞MDA-MB-231、MCF-7的影响,分析预测miR-96-5p与HOXA9的靶向作用关系。WB检测乳腺癌、癌旁组织、细胞HOXA9、Wnt/β-连环蛋白(β-catenin)信号通路蛋白表达,统计裸鼠成瘤体积、质量、肝转移灶数量,免疫组化检测肿瘤HOXA9、β-catenin表达情况。结果qRT-PCR检测结果显示,乳腺癌组织miR-96-5p水平升高(P<0.05),HOXA9水平降低(P<0.05)。miR-96-5p高表达与肿瘤大小≥3 cm、淋巴结转移阳性、TNM分期Ⅲ-Ⅳ期相关(P<0.05)。抑制miR-96-5p表达可降低MDA-MB-231、MCF-7细胞增殖、克隆形成、迁移、侵袭能力、Wnt-1、β-catenin、Cyclin D1表达水平、裸鼠肿瘤体积、质量、肝转移灶数量(P<0.05),提高HOXA9表达水平(P<0.05)。miR-96-5p靶向调控HOXA9表达,过表达HOXA9可逆转miR-96-5p对乳腺癌细胞恶性生物学行为的促进效果。结论miR-96-5p通过抑制HOXA9表达激活Wnt/β-catenin信号通路,促进乳腺癌发展。Objective To explore the mechanism of miR-96-5p targeted homeobox A9(HOXA9)promoting the proliferation and invasion of breast cancer cells.Methods Samples and complete clinical data of 60 cases of breast cancer and paracancerous tissues were resected from September 2022 to December 2022 were collected.Two pairs of differentially expressed miRNA in breast cancer and paracancerous tissues were analyzed by miRNA microarray.The expression of miR-96-5p in breast cancer,paracancerous tissues and breast cancer cell lines was detected by qRT-PCR,and the relationship between miR-96-5p expression and clinicopathological features was analyzed.The effects of miR-96-5p on breast cancer cells MDA-MB-231 and MCF-7 were detected by MTT,clone formation,EdU method,scratch,Transwell test and xenotransplantation in vivo.Analysis and prediction of the targeting relationship between miR-96-5p and HOXA9.Detection of HOXA9 and Wnt/β-catenin signal pathway protein expression in tumor,paracancerous tissues and breast cancer cells by WB.Statistics of tumor volume,mass and number of liver metastases in nude mice.The expression of HOXA9 andβ-catenin were detected by immunohistochemistry.Results The results of qRT-PCR detection showed that the level of miR-96-5p in breast cancer tissue increased and the level of HOXA9 decreased(P<0.05).The patients with high expression of miR-96-5p was associated with tumor size≥3 cm,positive lymph node metastasis and TNM stageⅢ-Ⅳ(P<0.05).Inhibition of miR-96-5p expression decreased the proliferation,clone formation,migration,invasion,Wnt-1,β-catenin,Cyclin D1 expression level,tumor volume,mass and number of liver metastases in nude mice(P<0.05),and increased HOXA9 expression level(P<0.05).miR-96-5p targeted regulated HOXA9 expression.Overexpression of HOXA9 can reverse the promoting effect of miR-96-5p on malignant biological behavior of breast cancer cells.Conclusion miR-96-5p activates the Wnt/β-catenin signaling pathway by inhibiting HOXA9 expression and promotes the development of breast cancer

关 键 词：miR-96-5p 同源盒基因A9 乳腺癌 增殖 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]