中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
5期
932-934
,共3页
田南南%孙国栋%郑名华%杨建新
田南南%孫國棟%鄭名華%楊建新
전남남%손국동%정명화%양건신
三阴性乳腺癌%微小RNA-10b%T淋巴瘤侵袭转移诱导因子1%侵袭
三陰性乳腺癌%微小RNA-10b%T淋巴瘤侵襲轉移誘導因子1%侵襲
삼음성유선암%미소RNA-10b%T림파류침습전이유도인자1%침습
Triple-negative breast cancer%microRNA-10b%T lymphoma invasion and metastasis inducing factor 1%Invasion
目的 观察微小RNA-10b(miR-10b)沉默对人三阴性乳腺癌MDA-MB-231细胞生物学行为的影响,并探讨其机制.方法 用人工合成的miR-10b inhibitor转染MDA-MB-231细胞,同时设空白细胞组和无义序列转染组为对照组,实时定量聚合酶链反应(Real-time PCR)检测转染后miR-10b的表达,以检验沉默效果.CCK-8法、流式细胞术、Transwell侵袭实验分析细胞增殖、凋亡、侵袭和迁移能力变化.Real-time PCR和Western blot法分别检测T淋巴细胞侵袭转移诱导因子1(TIAMl) mRNA和蛋白的表达.结果 与对照组比较,miR-10b inhibitor转染组的2-△△Ct值(0.26±0.01)明显下调(P＜0.01),表明成功沉默miR-10b的表达.miR-10b沉默后,48 h和72 h细胞增殖率分别为(409.10 ±8.38)％和(610.70±17.59)％,均明显上调(P＜0.01),增殖效应在48 h之后出现,并一直维持到72 h;早期和晚期细胞凋亡比例分别为1.77％和1.61％,均明显下调(P＜0.05);穿过基质膜的侵袭和迁移细胞数分别为(212.17±13.57)个和(322.67±8.62)个,均明显上调(P＜0.01).TIAM1基因在mRNA水平变化不大(P＞0.05),在蛋白水平表达上调(P＜0.05).结论 miR-10b沉默可以促进MDA-MB-231细胞的增殖、侵袭和迁移,并抑制凋亡,其机制可能与miR-10b沉默后在转录后水平上调TIAM1蛋白的表达有关.
目的 觀察微小RNA-10b(miR-10b)沉默對人三陰性乳腺癌MDA-MB-231細胞生物學行為的影響,併探討其機製.方法 用人工閤成的miR-10b inhibitor轉染MDA-MB-231細胞,同時設空白細胞組和無義序列轉染組為對照組,實時定量聚閤酶鏈反應(Real-time PCR)檢測轉染後miR-10b的錶達,以檢驗沉默效果.CCK-8法、流式細胞術、Transwell侵襲實驗分析細胞增殖、凋亡、侵襲和遷移能力變化.Real-time PCR和Western blot法分彆檢測T淋巴細胞侵襲轉移誘導因子1(TIAMl) mRNA和蛋白的錶達.結果 與對照組比較,miR-10b inhibitor轉染組的2-△△Ct值(0.26±0.01)明顯下調(P＜0.01),錶明成功沉默miR-10b的錶達.miR-10b沉默後,48 h和72 h細胞增殖率分彆為(409.10 ±8.38)％和(610.70±17.59)％,均明顯上調(P＜0.01),增殖效應在48 h之後齣現,併一直維持到72 h;早期和晚期細胞凋亡比例分彆為1.77％和1.61％,均明顯下調(P＜0.05);穿過基質膜的侵襲和遷移細胞數分彆為(212.17±13.57)箇和(322.67±8.62)箇,均明顯上調(P＜0.01).TIAM1基因在mRNA水平變化不大(P＞0.05),在蛋白水平錶達上調(P＜0.05).結論 miR-10b沉默可以促進MDA-MB-231細胞的增殖、侵襲和遷移,併抑製凋亡,其機製可能與miR-10b沉默後在轉錄後水平上調TIAM1蛋白的錶達有關.
목적 관찰미소RNA-10b(miR-10b)침묵대인삼음성유선암MDA-MB-231세포생물학행위적영향,병탐토기궤제.방법 용인공합성적miR-10b inhibitor전염MDA-MB-231세포,동시설공백세포조화무의서렬전염조위대조조,실시정량취합매련반응(Real-time PCR)검측전염후miR-10b적표체,이검험침묵효과.CCK-8법、류식세포술、Transwell침습실험분석세포증식、조망、침습화천이능력변화.Real-time PCR화Western blot법분별검측T림파세포침습전이유도인자1(TIAMl) mRNA화단백적표체.결과 여대조조비교,miR-10b inhibitor전염조적2-△△Ct치(0.26±0.01)명현하조(P＜0.01),표명성공침묵miR-10b적표체.miR-10b침묵후,48 h화72 h세포증식솔분별위(409.10 ±8.38)％화(610.70±17.59)％,균명현상조(P＜0.01),증식효응재48 h지후출현,병일직유지도72 h;조기화만기세포조망비례분별위1.77％화1.61％,균명현하조(P＜0.05);천과기질막적침습화천이세포수분별위(212.17±13.57)개화(322.67±8.62)개,균명현상조(P＜0.01).TIAM1기인재mRNA수평변화불대(P＞0.05),재단백수평표체상조(P＜0.05).결론 miR-10b침묵가이촉진MDA-MB-231세포적증식、침습화천이,병억제조망,기궤제가능여miR-10b침묵후재전록후수평상조TIAM1단백적표체유관.
Objective To investigate the eff~t of silencing microRNA-10b,on the biological behavior of hunan triple-negative breast cancer MDA-MB-231 cells and the mechanism Methods MDA-MB-231 cells were transfected with artificially synthesized miR-10b inhibitor.Glank cell group and nonsence sequence-transfected group were established as a control.Real-time quantitative polymerase chain reaction (Real-time PCR) was conducted to detect the expression of miR-10b after transfection.Cell counting kit-8 (CCK-8) assays,flow cytometry and Transwell assays were used to detect variations in cell proliferation,apoptosis,invasion and migration respectively.Real-time PCR and Western blotting were used to detect the expression of T lymphoma invasion and metastasis inducing factor 1 (TIAM1) mRNA and protein respec-tively.Results As compared with control group,the value of 2-△△Dt(0.26 ± 0.01) in miR-10b inhibitor-transfected group was significantly reduced (P ＜ 0.01),suggesting the expression of miR-10b was sup-pressed by miR-10b inhibitor successfully.The proliferation rate at 48 h [(409.10 ± 8.38) ％] and at 72 h [(610.70 ± 17.59) ％] was significantly increased (P ＜ 0.01) after silencing miR-10b.Proliferation effect occured at 48 h and continued to 72 h.Early apoptosis rate (1.77％) and late apoptosis rate (1.61％) were significantly reduced (P ＜0.05).The number of invasion (212.17 ± 13.57) and migration (322.67 ± 8.62) cells was significantly increased (P ＜ 0.01).TIAM1 gene had no significant changes at mRNA level (P ＞ 0.05),but the protein expression was up-regurated (P ＜ 0.05).Conclusion Silencing miR-10b could promote proliferation,invasion and migration,and inhibit apoptosis of MDA-MB-231 cells,which is probably related to the positive regulation of TIAM1 protein at the post-transcriptional 1 evel.
