title: Inhibition of microRNA-196a might reverse cisplatin resistance of A549/DDP non-small-cell lung cancer cell line.
abstract: We aimed to explore the possible mechanism of microRNA-196a (miR-196a) inhibition and reversion of drug resistance to cisplatin (DDP) of the A549/DDP non-small-cell lung cancer (NSCLC) cell line. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect expression differences of miR-196a in the drug-resistant A549/DDP NLCLC cell line and the parental A549 cell line, and expressions of miR-196a in the A549/DDP NLCLC cell line transfected with miR-196a inhibitor (anti-miR-196a group) and the miR-196a negative control (miR-NC) group and blank group (without transfection). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was applied in examining the cell viability of A549/DDP cell line before and after transfection. Clonogenic assay was used to detect cell proliferation ability. Flow cytometry was applied in detecting apoptosis rate of assayed tumor cell and rhodamine-123 changes in cells. Western blot was applied in detecting proteins of drug-resistant related gene in A549/DDP cell line. Significantly higher expression of miR-196a was detected in the drug-resistant A549/DDP cell line than that in the parental A549 cell line (P < 0.05). However, miR-196a expression in the anti-miR-196a group decreased obviously compared to that in the blank group and the miR-NC group (both P < 0.05); The value of IC50 in the anti-miR-196a group showed remarkably lower than that in the blank group and the miR-NC group (both P < 0.05); Rh-123 absorbing ability in the anti-miR-196a group increased 2.51 times and 2.49 times respectively compared to that in the blank group and the miR-NC group (both P < 0.05). No statistical differences in the apoptosis rate of A549/DDP cell line in the early stage were found among the three groups (all P > 0.05), but the late-stage apoptosis rate in the anti-miR-196a group was significantly higher than that in the blank group and the miR-NC group (both P < 0.05); The expressions of human multidrug resistance 1 (MDR1), multidrug resistance protein 1 (MRP1), excision repair cross-complementation 1 (ERCC1), survivin, and B cell lymphoma 2 (Bcl-2) decreased significantly while RhoE increased significantly in the anti-miR-196a group than the blank group and the miR-NC group (all P < 0.05). Inhibition of miR-196a could reverse cisplatin resistance of A549/DDP cell lines, which might relate with inhibition of drug efflux, down-regulation of drug-resistant protein expression, cell apoptosis, and cell proliferation suppression.
authors.affiliation: Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Jian-Huang
authors.initials: JH
authors.lastname: Li
authors.affiliation: Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Ning
authors.initials: N
authors.lastname: Luo
authors.affiliation: Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Mei-Zuo
authors.initials: MZ
authors.lastname: Zhong
authors.affiliation: Research Center of Carcinogenesis and Targeted Therapy, Xiangya Hospital, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Xiao
authors.affiliation: School of Information Science and Engineering, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Jian-Xin
authors.initials: JX
authors.lastname: Wang
authors.affiliation: Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Xiao-Yi
authors.initials: XY
authors.lastname: Yao
authors.affiliation: Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410000, People's Republic of China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Department of Medical Oncology, the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Tongzipo Road No. 283, Yuelu District, Changsha, 410000, People's Republic of China. caojun0939@163.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Cao
mesh_terms: D000072283:A549 Cells; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000235:genetics; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D002945:Cisplatin / Q000493:pharmacokinetics*; D015536:Down-Regulation / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D014162:Transfection / Q000379:methods
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D051034:Inhibitor of Apoptosis Proteins; C542543:MIRN196 microRNA, human; D035683:MicroRNAs; D002945:Cisplatin
keywords: A549/DDP cell lines; Cisplatin resistance; MicroRNA-196a; NSCLC