肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2014年
10期
649-653
,共5页
何龙%杨宏伟%张岩%王军%刘龙%范连慧
何龍%楊宏偉%張巖%王軍%劉龍%範連慧
하룡%양굉위%장암%왕군%류룡%범련혜
癌,肾细胞%miR-218%凋亡%增殖%抑癌基因
癌,腎細胞%miR-218%凋亡%增殖%抑癌基因
암,신세포%miR-218%조망%증식%억암기인
Carcinoma,renal cell%miR-218%Apoptosis%Proliferation%Anti-oncogene
目的 通过调控miR-218在肾癌细胞株中的表达水平,验证miR-218对肾细胞癌细胞生物学功能的影响.方法 将pcDNA3.1-miR-218稳定转染至肾癌细胞株A498、769-P中,检测肾癌细胞株中miR-218表达水平的变化,采用CCK8、Transwell小室、Annxin V-FITC、生长曲线、克隆实验等,体外验证转染后的癌细胞株在miR-218调控下的细胞活性、侵袭能力、凋亡情况、增殖能力的变化.结果 A498、769-P细胞转染后miR-218相对表达量分别为1.99、1.64,较转染前表达量(1.00)增高,差异均有统计学意义(t=60.82、10.89,均P＜0.000 1).A498、769-P细胞转染miR-218表达质粒组的细胞活性分别为0.90±0.10、0.68±0.06,低于对照组的1.39±0.14、1.24±0.08,差异均有统计学意义(t=15.02、31.69,均P＜0.000 1).Transwell检测结果显示,A498、769-P细胞转染miR-218表达质粒组的细胞侵袭能力较对照组减弱(t=15.78、18.80,均P＜0.000 1).AnnxinV-FITC检测结果显示,A498、769-P细胞株对照组和转染组凋亡细胞比例分别为0.25％、45.77％和0.11％、45.57％.转染组细胞增殖能力较对照组减弱(均P＜ 0.000 1).结论 体外实验证实miR-218表达上调能够抑制肾癌细胞的生长.
目的 通過調控miR-218在腎癌細胞株中的錶達水平,驗證miR-218對腎細胞癌細胞生物學功能的影響.方法 將pcDNA3.1-miR-218穩定轉染至腎癌細胞株A498、769-P中,檢測腎癌細胞株中miR-218錶達水平的變化,採用CCK8、Transwell小室、Annxin V-FITC、生長麯線、剋隆實驗等,體外驗證轉染後的癌細胞株在miR-218調控下的細胞活性、侵襲能力、凋亡情況、增殖能力的變化.結果 A498、769-P細胞轉染後miR-218相對錶達量分彆為1.99、1.64,較轉染前錶達量(1.00)增高,差異均有統計學意義(t=60.82、10.89,均P＜0.000 1).A498、769-P細胞轉染miR-218錶達質粒組的細胞活性分彆為0.90±0.10、0.68±0.06,低于對照組的1.39±0.14、1.24±0.08,差異均有統計學意義(t=15.02、31.69,均P＜0.000 1).Transwell檢測結果顯示,A498、769-P細胞轉染miR-218錶達質粒組的細胞侵襲能力較對照組減弱(t=15.78、18.80,均P＜0.000 1).AnnxinV-FITC檢測結果顯示,A498、769-P細胞株對照組和轉染組凋亡細胞比例分彆為0.25％、45.77％和0.11％、45.57％.轉染組細胞增殖能力較對照組減弱(均P＜ 0.000 1).結論 體外實驗證實miR-218錶達上調能夠抑製腎癌細胞的生長.
목적 통과조공miR-218재신암세포주중적표체수평,험증miR-218대신세포암세포생물학공능적영향.방법 장pcDNA3.1-miR-218은정전염지신암세포주A498、769-P중,검측신암세포주중miR-218표체수평적변화,채용CCK8、Transwell소실、Annxin V-FITC、생장곡선、극륭실험등,체외험증전염후적암세포주재miR-218조공하적세포활성、침습능력、조망정황、증식능력적변화.결과 A498、769-P세포전염후miR-218상대표체량분별위1.99、1.64,교전염전표체량(1.00)증고,차이균유통계학의의(t=60.82、10.89,균P＜0.000 1).A498、769-P세포전염miR-218표체질립조적세포활성분별위0.90±0.10、0.68±0.06,저우대조조적1.39±0.14、1.24±0.08,차이균유통계학의의(t=15.02、31.69,균P＜0.000 1).Transwell검측결과현시,A498、769-P세포전염miR-218표체질립조적세포침습능력교대조조감약(t=15.78、18.80,균P＜0.000 1).AnnxinV-FITC검측결과현시,A498、769-P세포주대조조화전염조조망세포비례분별위0.25％、45.77％화0.11％、45.57％.전염조세포증식능력교대조조감약(균P＜ 0.000 1).결론 체외실험증실miR-218표체상조능구억제신암세포적생장.
Objective To explore the effects of miR-218 on biological functions of renal cell carcinoma cell line through regulating the expression levels of miR-218.Methods The pcDNA3.1-miR-218 was transfected into renal carcinoma cell line A498 and 769-P and the expression of miR-218 was detected.The cell activity,invasion,apoptosis and proliferation of transfected renal carcinoma cell line were analyzed in vitro.Results After plasmid transfected A498/769-P renal carcinoma cell line,the expression level of miR-218 (1.99,1.64) was significantly higher than that of the control group (1.00) (t =60.82,10.89,P ＜ 0.000 1) The cell viability (0.90±0.10,0.68±0.06) was lower than that of the control group (1.39±0.14,1.24±0.08) by CCK8 experiment (t =15.02,31.69,P ＜ 0.000 1).The cell invasion was lower than that of control group by Transwell experiment (t =15.78,18.80,P ＜ 0.000 1).The apoptosis ratio was higher by AnnxinV-FITC experiment,the apoptosis ration of control group and transfected group were respectively 0.25 ％,45.77 ％ in A498 cell line and 0.11％,45.57 ％ in 769-P cell line.The proliferation was lower than that of the control group (P ＜ 0.000 1).Conclusion Up-regulation of miR-218 expression can inhibit the growth of renal cell carcinoma cell line in vitro.
