miR-221-3p靶向沉默PARP1对三阴乳腺癌细胞的化疗药物敏感性的影响  

miR-221-3p targeted silencing of PARP1 increases the sensitivity of triple negative breast cancer cells to chemotherapy drug

作　　者：马楠[1] 齐宝文[2] 徐蓉[3] MA Nan;QI Bao-wen;XU Rong(Department of Breast and Thyroid Surgery,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 83000;不详)

机构地区：[1]新疆维吾尔自治区人民医院乳腺甲状腺科,新疆乌鲁木齐830001 [2]新疆维吾尔自治区人民医院超声科,新疆乌鲁木齐830001 [3]新疆维吾尔自治区人民医院肿瘤科,新疆乌鲁木齐830001

出　　处：《广东医学》2023年第8期925-933,共9页Guangdong Medical Journal

基　　金：新疆维吾尔自治区自然科学基金资助项目(2020D01C117)。

摘　　要：目的分析miR-221-3p通过PARP1调节三阴乳腺癌(TNBC)细胞化疗耐药的分子机制。方法收集行肿瘤切除术的TNBC患者标本,通过实时荧光定量PCR和Western blot检测癌旁组织、TNBC组织、TNBC化疗耐药组织和TNBC化疗敏感组织中miR-221-3p及其PARP1表达,取对数期MDA-MB-231细胞,将添加0.1μg/mL ADM的培养基加入细胞内,培养获得耐药的MDA-MB-231/ADM细胞。miR-221-3p inhibitor与inhibitor NC分别转染MDA-MB-231/ADM细胞,CCK-8法检测细胞药物敏感性,流式细胞仪分析细胞凋亡率,Western blot实验分析肿瘤细胞内Bax、Caspase-3、Caspase-9和Bcl-2蛋白的表达情况。TargetScan数据库预测miR-221-3p的潜在靶点,双荧光素酶报告基因实验分析miR-221-3p与PARP1之间的关系,Western blot实验检测下调miR-221-3p表达后PARP1、BRD7蛋白表达情况。PARP1 siRNA与siRNA NC分别转染MDA-MB-231/ADM细胞,分别利用CCK-8、流式细胞仪和Western blot检测细胞药物敏感性和凋亡情况。最后分析上调miR-221-3p通过PARP1对MDA-MB-231/ADM细胞药物敏感性和凋亡的影响。结果与癌旁组织相比,TNBC组织中miR-221-3p表达下调(P<0.01),PARP1表达上调(P<0.01);与TNBC化疗敏感组织相比,TNBC化疗耐药组织中miR-221-3p表达下调(P<0.01),PARP1表达上调(P<0.01)。下调了miR-221-3p表达抑制了MDA-MB-231/ADM细胞的药物敏感性,细胞凋亡率明显降低。miR-221-3p靶向PARP1,下调miR-221-3p促进了PARP1蛋白表达,而抑制了BRD7蛋白表达,PARP1 siRNA转染MDA-MB-231/ADM细胞后促进了BRD7蛋白表达和细胞凋亡,上调了细胞药物敏感性。而过表达miR-221-3p通过PARP1上调了细胞凋亡和药物敏感性。结论miR-221-3p靶向沉默PARP1增加TNBC细胞对化疗药物的敏感性。Objective To analyze the molecular mechanism of miR-221-3p regulating chemotherapy resistance in triple-negative breast cancer(TNBC)cells via PARP1.Methods Tumor specimens from patients who underwent tumor resection for TNBC were collected.Real-time quantitative PCR and Western blot were used to detect the expression levels of miR-221-3p and PARP1 in adjacent tissues,TNBC tissues,TNBC chemoresistant tissues,and TNBC chemosensitive tissues.Logarithmic phase MDA-MB-231 cells were used to establish ADM-resistant MDA-MB-231/ADM cells by adding 0.1μg/mL ADM to the culture medium.MDA-MB-231/ADM cells were transfected with miR-221-3p inhibitor or inhibitor NC,and cell drug sensitivity was detected using the CCK-8 assay.Flow cytometry was used to analyze the apoptosis rate,and Western blot experiments were performed to analyze the expression of Bax,Caspase-3,Caspase-9,and Bcl-2 proteins in tumor cells.TargetScan database was used to predict potential targets of miR-221-3p,and dual-luciferase reporter gene experiments were performed to analyze the relationship between miR-221-3p and PARP1.Western blot was used to detect the expression of PARP1 and BRD7 proteins after downregulation of miR-221-3p.MDA-MB-231/ADM cells were transfected with PARP1 siRNA or siRNA NC,and drug sensitivity and apoptosis were detected using CCK-8,flow cytometry,and Western blot,respectively.Finally,the impact of upregulating miR-221-3p on drug sensitivity and apoptosis in MDA-MB-231/ADM cells through PARP1 was analyzed.Results Compared with adjacent tissues,miR-221-3p expression was downregulated(P<0.01),and PARP1 expression was upregulated in TNBC tissues(P<0.01).Compared with TNBC chemosensitive tissues,miR-221-3p expression was downregulated(P<0.01),and PARP1 expression was upregulated in TNBC chemoresistant tissues(P<0.01).Downregulation of miR-221-3p inhibited drug sensitivity and significantly reduced the apoptosis rate of MDA-MB-231/ADM cells.MiR-221-3p targeted PARP1,downregulated miR-221-3p promoted PARP1 protein expression and inhibited

关 键 词：三阴乳腺癌 miR-221-3p PARP1 化疗敏感性 

分 类 号：R737.9[医药卫生—肿瘤] R392[医药卫生—临床医学]