CAJ | 학술논문

目的：观察前列腺癌PC-3细胞中微小RNA（microRNA，miRNA）-221和miRNA-222对细胞增殖和迁移的作用，以及对沉默信息调节因子1（SIRT1）表达的影响。方法设立无处理细胞组（简称control组）、转染空载质粒pEX-5组（简称pEX-5组）、转染miRNA-221反义抑制质粒组（简称miRNA-221抑制组）、转染miRNA-222反义抑制质粒组（简称miRNA-222抑制组），应用细胞计数试剂盒（CCK-8法）检测miRNA-221和miRNA-222对前列腺癌细胞增殖的影响；采用细胞划痕修复实验检测miRNA-221和miRNA-222对细胞迁移能力的影响；采用免疫印迹法和荧光定量聚合酶链反应（PCR）检测SIRT1在蛋白水平和mRNA水平的变化；采用miRNAanda靶标进行预测分析。结果在成功抑制了PC-3细胞中miRNA-221或miRNA-222的表达后，连续7 d检测细胞活性，miRNA-221抑制组和miRNA-222抑制组的细胞活性（A450 nm值）较pEX-5组降低1．5～2．0倍，自第3天起两组之间即有明显差异（t＝6．7，P＜0．01；t＝5．3，P＜0．01）；划痕实验显示，miRNA-221抑制组和miRNA-222抑制组的迁移能力明显低于pEX-5组。miRNA-221抑制组、miRNA-222抑制组中SIRT1蛋白的相对表达量分别为（0．26±0．021）、（0．21±0．0058），与pEX-5组（0．14±0．017）比较差异均有统计学意义（t值分别为6．3、6．4，P均＜0．05）；而SIRT1 mRNA的表达水平3组之间差异无统计学意义（P＞0．05）。结论 miRNA-221和miRNA-222能促进前列腺癌PC-3细胞的增殖与迁移能力，影响SIRT1蛋白的表达。
목적：관찰전렬선암PC-3세포중미소RNA（microRNA，miRNA）-221화miRNA-222대세포증식화천이적작용，이급대침묵신식조절인자1（SIRT1）표체적영향。방법설립무처리세포조（간칭control조）、전염공재질립pEX-5조（간칭pEX-5조）、전염miRNA-221반의억제질립조（간칭miRNA-221억제조）、전염miRNA-222반의억제질립조（간칭miRNA-222억제조），응용세포계수시제합（CCK-8법）검측miRNA-221화miRNA-222대전렬선암세포증식적영향；채용세포화흔수복실험검측miRNA-221화miRNA-222대세포천이능력적영향；채용면역인적법화형광정량취합매련반응（PCR）검측SIRT1재단백수평화mRNA수평적변화；채용miRNAanda파표진행예측분석。결과재성공억제료PC-3세포중miRNA-221혹miRNA-222적표체후，련속7 d검측세포활성，miRNA-221억제조화miRNA-222억제조적세포활성（A450 nm치）교pEX-5조강저1．5～2．0배，자제3천기량조지간즉유명현차이（t＝6．7，P＜0．01；t＝5．3，P＜0．01）；화흔실험현시，miRNA-221억제조화miRNA-222억제조적천이능력명현저우pEX-5조。miRNA-221억제조、miRNA-222억제조중SIRT1단백적상대표체량분별위（0．26±0．021）、（0．21±0．0058），여pEX-5조（0．14±0．017）비교차이균유통계학의의（t치분별위6．3、6．4，P균＜0．05）；이SIRT1 mRNA적표체수평3조지간차이무통계학의의（P＞0．05）。결론 miRNA-221화miRNA-222능촉진전렬선암PC-3세포적증식여천이능력，영향SIRT1단백적표체。
Objective To investigate the roles of micro RNA(miRNA)-221 and miRNA-222 on the proliferation and migration of prostate cancer cell PC-3 and the influence on silent information regulator 1 (SIRT1 ).Methods The groups were arranged as cells without treatment (control group),cells transfected with pEX-5 empty plasmid (pEX-5 group), cells transfected with miRNA-221 inhibitor (miRNA-221 inhibition group)and cells transfected with miRNA-222 inhibitor (miRNA-222 inhibition group).The influence of miRNA-221 and miRNA-222 on prostate cancer cell proliferation was determined by CCK-8,and the migration ability was determined by wound healing test.The protein and mRNA levels of SIRT1 were determined by Western blotting and fluorescence quantitation polymerase chain reaction (PCR),respectively. The prediction analysis was performed by miRNAanda target.Results After the expression inhibition of miRNA-221 or miRNA-222 in cell PC-3,the cell activities were determined for 7 d.The activities (A450 mm)in miRNA-221 and miRNA-222 inhibition groups were down-regulated about 1.5-2.0 times compared with that in pEX-5 group,and there were significant differences between the 2 groups from the 3rd d(t=6.7,P<0.01;t=5.3,P<0.01).The wound healing test showed that the migration abilities of miRNA-221 inhibition group and miRNA-222 inhibition group decreased than that of pEX-5 group.Moreover,the expression levels of SIRT1 protein in miRNA-221 inhibition group(0.26 ±0.021)and miRNA-222 inhibition group(0.21 ±0.005 8)were up-regulated compared with that in pEX-5 group(0.14 ±0.017),and there were statistical significances (t=6.3 and 6.4,P<0.05 ).The SIRT1 mRNA expression levels had no statistical significance among the 3 groups(P>0.05).Conclusions The miRNA-221 and miRNA-222 promote the proliferation and migration of prostate cancer cell PC-3 and have the influence on SIRT1 expression in protein level.