CAJ | 학술논문

目的探讨转染miRNA-30a-5p对肝癌细胞生物学行为的影响. 方法将miRNA-30a-5p模拟物、miRNA-30a-5p抑制物瞬时转染入肝细胞肝癌细胞株SMCC-7721,应用实时荧光定量PCR检测正常肝细胞株L02及肝癌细胞株SMCC-7721转染后的miR-30a-5p的mRNA表达情况,CCK-8法检测细胞增殖能力,克隆形成实验检测集落形成情况,流式细胞术检测细胞凋亡及各组细胞周期分布的差异,Transwell小室检测各组细胞体外侵袭转移能力,建立BALB/c-nu裸小鼠肝癌模型并观察miRNA-30a-5p对肿瘤生长的影响. 结果实时荧光定量PCR显示,与未转染组及正常肝细胞株L02相比,肝癌细胞株SMCC-7721在转染miRNA-30a-5p模拟物后,mRNA表达明显上调(P＜0.01),而转染miRNA-30a-5p抑制物的mRNA表达明显受抑(P＜0.01),差异有统计学意义.肝细胞肝癌细胞株SMCC-7721在转染miRNA-30a-5p模拟物后,细胞活性、克隆形成能力、迁移和侵袭能力与miRNA-30a-5p抑制物转染组、未转染组及正常肝细胞株L02相比,相对减弱(P＜0.05),miRNA-30a-5p模拟物转染组凋亡率,高于miRNA-30a-5p抑制物转染组、未转染组及正常肝细胞株L02 (P＜0.05),细胞周期出现S期阻滞.裸鼠肝癌模型中,实验组裸鼠瘤体质量及体积明显小于空载体对照组和空白对照组(P＜ 0.05).结论上调miR-30a-5p表达可明显抑制肝细胞肝癌细胞株SMCC-7721的增殖,促进其凋亡,抑制其迁移、侵袭能力,并且抑制裸小鼠肝癌模型肿瘤的生长.
목적 탐토전염miRNA-30a-5p대간암세포생물학행위적영향. 방법 장miRNA-30a-5p모의물、miRNA-30a-5p억제물순시전염입간세포간암세포주SMCC-7721,응용실시형광정량PCR검측정상간세포주L02급간암세포주SMCC-7721전염후적miR-30a-5p적mRNA표체정황,CCK-8법검측세포증식능력,극륭형성실험검측집락형성정황,류식세포술검측세포조망급각조세포주기분포적차이,Transwell소실검측각조세포체외침습전이능력,건립BALB/c-nu라소서간암모형병관찰miRNA-30a-5p대종류생장적영향. 결과 실시형광정량PCR현시,여미전염조급정상간세포주L02상비,간암세포주SMCC-7721재전염miRNA-30a-5p모의물후,mRNA표체명현상조(P＜0.01),이전염miRNA-30a-5p억제물적mRNA표체명현수억(P＜0.01),차이유통계학의의.간세포간암세포주SMCC-7721재전염miRNA-30a-5p모의물후,세포활성、극륭형성능력、천이화침습능력여miRNA-30a-5p억제물전염조、미전염조급정상간세포주L02상비,상대감약(P＜0.05),miRNA-30a-5p모의물전염조조망솔,고우miRNA-30a-5p억제물전염조、미전염조급정상간세포주L02 (P＜0.05),세포주기출현S기조체.라서간암모형중,실험조라서류체질량급체적명현소우공재체대조조화공백대조조(P＜ 0.05).결론 상조miR-30a-5p표체가명현억제간세포간암세포주SMCC-7721적증식,촉진기조망,억제기천이、침습능력,병차억제라소서간암모형종류적생장.
Objective To explore the effect ofmicroRNA-30a-5p (miRNA-30a-5p) on the biological behavior of human hepatoma cells.Methods The liver cancer cell line SMCC-7721 cells and the normal liver cell line L02 cells (control) were transiently transfected with miRNA-30a-5p mimics and an miRNA-30a-5p inhibitor by Lipofectamine 2000 (Life Technologies).miR-30a-5p mRNA expression was detected by quantitative real-time (q)PCR.Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8) assay and apoptosis was assessed by flow cytometry.Invasion and migration were measured by transwell chamber assays.The SMCC-7721 cells was injected subcutaneously into nude mice to establish a tumor animal model.Results The SMCC-7721 cells transfected with miRNA-30a-5p mimics showed significantly higher miRNA-30a-5p mRNA expression than the non-transfected SMCC-7721 cells and the transfected control L02 cells (P ＜ 0.01).The miRNA-30a-5p mRNA expression was significantly lower in the SMCC-7721 cells transfected with the miRNA-30a-5p inhibitor than the non-transfected SMCC-7721cells the control L02 cells (P ＜ 0.01).The overexpression of miRNA-30a-5p inhibited the viability,colony formation rate,and invasion and migration abilities,as shown in the cells transfected with the miRNA-30a-5p mimics (P ＜ 0.05); in addition,the miRNA-30a-5p promoted proliferation of cells (P ＜ 0.05),as shown by more S phase cells detected by flow cytometry.SMCC-7122 cells transfected with miRNA-30a-5p mimics produced tumors with significantly higher average weight than tumors produced by SMCC-7122 cells that were untransfected or transfected with empty vector (bothP ＜ 0.01).Conclusion Overexpression ofmiR-30a-5p had an inhibitory effect on cell proliferation,induced apoptosis,increased the number of cells in S phase,and markedly inhibited invasion and migration of SMCC-7721 HCC cells in vitro and in vivo.