中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2014年
4期
250-256
,共7页
食管肿瘤%癌,鳞状细胞%EC9706细胞%T细胞淋巴瘤侵袭转移诱导因子1%RNA,小分子干扰%细胞增殖%细胞周期%细胞凋亡
食管腫瘤%癌,鱗狀細胞%EC9706細胞%T細胞淋巴瘤侵襲轉移誘導因子1%RNA,小分子榦擾%細胞增殖%細胞週期%細胞凋亡
식관종류%암,린상세포%EC9706세포%T세포림파류침습전이유도인자1%RNA,소분자간우%세포증식%세포주기%세포조망
Esophageal neoplasms%Carcinoma,squamous cell%EC9706 cells%T-cell lymphoma invasion and metastasis 1%RNA,small interfering%Cell proliferation%Cell cycle%Apoptosis
目的 探讨小干扰RNA(siRNA)下调T细胞淋巴瘤侵袭转移诱导因子1(Tiam1)表达对食管鳞癌EC9706细胞的影响.方法 利用Tiam1 siRNA转染食管鳞癌EC9706细胞,采用实时荧光定量PCR和Western blot检测转染后Tiam1 mRNA和蛋白的表达,CCK-8试剂盒分析转染后细胞的增殖能力,流式细胞术检测细胞周期和细胞凋亡情况,Western b1ot检测细胞周期相关蛋白和细胞凋亡相关蛋白的表达.结果 未处理组和对照siRNA组EC9706细胞中Tiam1 mRNA的相对表达水平分别为1.00和0.95±0.02,差异无统计学意义(P＞0.05).Tiam1 siRNA转染24、48、72 h后,EC9706细胞中Tiam1 mRNA的相对表达水平分别为0.30 ±0.04、0.09 ±0.01和0.09 ±0.006,与未处理组比较差异均有统计学意义(均P＜0.05).转染Tiam1 siRNA 24 h后,EC9706细胞中Tiam1蛋白的表达水平(0.11 ±0.02)低于未处理组(0.44±0.05)和对照siRNA组(0.44±0.04,均P＜0.05).Tiam1 siRNA组、未处理组和对照siRNA组Go/G1期细胞所占比例分别为(54.48±2.14)％、(40.69±1.85)％和(41.78±1.31)％,S期细胞所占比例分别为(27.18±1.65)％、(32.32±1.15)％和(30.35±1.09)％,差异均有统计学意义(均P＜0.01).未处理组、对照siRNA组和Tiam1siRNA组中cyclin D1蛋白的表达水平分别为0.43 ±0.02、0.41 ±0.01和0.11±0.02,p27蛋白的表达水平分别为0.10±0.01、0.09 ±0.02和0.20±0.02,差异均有统计学意义(均P＜0.05).未处理组、对照siRNA组和Tiam1 siRNA组中早期凋亡细胞所占比例分别为(10±0.9)％、(10±0.5)％和(27±0.7)％,差异有统计学意义(P＜0.01).未处理组、对照siRNA组和Tiam1 siRNA组EC9706细胞中Mcl-1蛋白的表达水平分别为0.47 ±0.12、0.48 ±0.13和0.16±0.02,Bcl-2蛋白的表达水平分别为0.49 ±0.08、0.50 ±0.05和0.04 ±0.03,caspase-3的活性分别为2.3±0.09、2.3±0.10和16.0±1.50,caspase-9的活性分别为2.3 ±0.08、2.3 ±0.11和14.5 ±0.9,差异均有统计学意义(均P＜0.05).结论 Tiam1 siRNA能明显抑制食管鳞癌EC9706细胞的增殖、诱导细胞周期静止和细胞凋亡的发生,这与细胞周期基因(cyclin D1和p27)和细胞凋亡基因(Mcl-1、Bcl-1、caspase-3和caspase-9)的变化密切相关.
目的 探討小榦擾RNA(siRNA)下調T細胞淋巴瘤侵襲轉移誘導因子1(Tiam1)錶達對食管鱗癌EC9706細胞的影響.方法 利用Tiam1 siRNA轉染食管鱗癌EC9706細胞,採用實時熒光定量PCR和Western blot檢測轉染後Tiam1 mRNA和蛋白的錶達,CCK-8試劑盒分析轉染後細胞的增殖能力,流式細胞術檢測細胞週期和細胞凋亡情況,Western b1ot檢測細胞週期相關蛋白和細胞凋亡相關蛋白的錶達.結果 未處理組和對照siRNA組EC9706細胞中Tiam1 mRNA的相對錶達水平分彆為1.00和0.95±0.02,差異無統計學意義(P＞0.05).Tiam1 siRNA轉染24、48、72 h後,EC9706細胞中Tiam1 mRNA的相對錶達水平分彆為0.30 ±0.04、0.09 ±0.01和0.09 ±0.006,與未處理組比較差異均有統計學意義(均P＜0.05).轉染Tiam1 siRNA 24 h後,EC9706細胞中Tiam1蛋白的錶達水平(0.11 ±0.02)低于未處理組(0.44±0.05)和對照siRNA組(0.44±0.04,均P＜0.05).Tiam1 siRNA組、未處理組和對照siRNA組Go/G1期細胞所佔比例分彆為(54.48±2.14)％、(40.69±1.85)％和(41.78±1.31)％,S期細胞所佔比例分彆為(27.18±1.65)％、(32.32±1.15)％和(30.35±1.09)％,差異均有統計學意義(均P＜0.01).未處理組、對照siRNA組和Tiam1siRNA組中cyclin D1蛋白的錶達水平分彆為0.43 ±0.02、0.41 ±0.01和0.11±0.02,p27蛋白的錶達水平分彆為0.10±0.01、0.09 ±0.02和0.20±0.02,差異均有統計學意義(均P＜0.05).未處理組、對照siRNA組和Tiam1 siRNA組中早期凋亡細胞所佔比例分彆為(10±0.9)％、(10±0.5)％和(27±0.7)％,差異有統計學意義(P＜0.01).未處理組、對照siRNA組和Tiam1 siRNA組EC9706細胞中Mcl-1蛋白的錶達水平分彆為0.47 ±0.12、0.48 ±0.13和0.16±0.02,Bcl-2蛋白的錶達水平分彆為0.49 ±0.08、0.50 ±0.05和0.04 ±0.03,caspase-3的活性分彆為2.3±0.09、2.3±0.10和16.0±1.50,caspase-9的活性分彆為2.3 ±0.08、2.3 ±0.11和14.5 ±0.9,差異均有統計學意義(均P＜0.05).結論 Tiam1 siRNA能明顯抑製食管鱗癌EC9706細胞的增殖、誘導細胞週期靜止和細胞凋亡的髮生,這與細胞週期基因(cyclin D1和p27)和細胞凋亡基因(Mcl-1、Bcl-1、caspase-3和caspase-9)的變化密切相關.
목적 탐토소간우RNA(siRNA)하조T세포림파류침습전이유도인자1(Tiam1)표체대식관린암EC9706세포적영향.방법 이용Tiam1 siRNA전염식관린암EC9706세포,채용실시형광정량PCR화Western blot검측전염후Tiam1 mRNA화단백적표체,CCK-8시제합분석전염후세포적증식능력,류식세포술검측세포주기화세포조망정황,Western b1ot검측세포주기상관단백화세포조망상관단백적표체.결과 미처리조화대조siRNA조EC9706세포중Tiam1 mRNA적상대표체수평분별위1.00화0.95±0.02,차이무통계학의의(P＞0.05).Tiam1 siRNA전염24、48、72 h후,EC9706세포중Tiam1 mRNA적상대표체수평분별위0.30 ±0.04、0.09 ±0.01화0.09 ±0.006,여미처리조비교차이균유통계학의의(균P＜0.05).전염Tiam1 siRNA 24 h후,EC9706세포중Tiam1단백적표체수평(0.11 ±0.02)저우미처리조(0.44±0.05)화대조siRNA조(0.44±0.04,균P＜0.05).Tiam1 siRNA조、미처리조화대조siRNA조Go/G1기세포소점비례분별위(54.48±2.14)％、(40.69±1.85)％화(41.78±1.31)％,S기세포소점비례분별위(27.18±1.65)％、(32.32±1.15)％화(30.35±1.09)％,차이균유통계학의의(균P＜0.01).미처리조、대조siRNA조화Tiam1siRNA조중cyclin D1단백적표체수평분별위0.43 ±0.02、0.41 ±0.01화0.11±0.02,p27단백적표체수평분별위0.10±0.01、0.09 ±0.02화0.20±0.02,차이균유통계학의의(균P＜0.05).미처리조、대조siRNA조화Tiam1 siRNA조중조기조망세포소점비례분별위(10±0.9)％、(10±0.5)％화(27±0.7)％,차이유통계학의의(P＜0.01).미처리조、대조siRNA조화Tiam1 siRNA조EC9706세포중Mcl-1단백적표체수평분별위0.47 ±0.12、0.48 ±0.13화0.16±0.02,Bcl-2단백적표체수평분별위0.49 ±0.08、0.50 ±0.05화0.04 ±0.03,caspase-3적활성분별위2.3±0.09、2.3±0.10화16.0±1.50,caspase-9적활성분별위2.3 ±0.08、2.3 ±0.11화14.5 ±0.9,차이균유통계학의의(균P＜0.05).결론 Tiam1 siRNA능명현억제식관린암EC9706세포적증식、유도세포주기정지화세포조망적발생,저여세포주기기인(cyclin D1화p27)화세포조망기인(Mcl-1、Bcl-1、caspase-3화caspase-9)적변화밀절상관.
Objective To explore the effect of downregulation of Tiam1 by siRNA on the esophageal squamous cell carcinoma (ESCC) EC9706 cells,and provide theoretical basis for gene therapy of ESCC using Tiam1 as a molecular target.Methods Tiaml siRNA was transfected into EC9706 cells,and expression changes of Tiam1 mRNA and protein after transfection were detected by quantitative real-time PCR and Western blotting.Cell proliferation was analyzed using CCK-8 kit.Cell cycle and apoptosis of the EC9706 cells were assessed by flow cytometry.Cell cycle-related proteins and cell apoptosis-associated proteins were analyzed by Western blotting.Results Compared with the untreated group and control siRNA group,the relative expression levels of Tiam1 mRNA (1.00 and 0.11 ± 0.02) were not significantly different (P ＞0.05).The relative expression levels of Tiam1 mRNA in the Tiam1 siRNA group at 24,48 and 72 h after transfection were 0.30 ± 0.04,0.09 ± 0.01 and 0.09 ± 0.006,respectively,significantly lower than that of the untreated group (P ＜ 0.05 for all).The expression level of Tiam1 protein at 24 h after Tiam1 siRNA transfection in the EC9706 cells was 0.11 ±0.02,significantly lower than that in the un-treated group (0.44 ± 0.05) and control siRNA group (0.44 ± 0.04,P ＜ 0.05 for all).The percentages of G0/G1 cells in the Tiam1 siRNA group,untreated group and control siRNA group were (54.48 ± 2.14)％,(40.69 ± 1.85) ％ and (41.78 ± 1.31) ％,respectively (P ＜ 0.01).The percentages of S phase cells in the Tiam1 siRNA group,untreated group and control siRNA group were (27.18 ± 1.65) ％,(32.32 ± 1.15) ％ and (30.35 ± 1.09)％,respectively (P ＜ 0.01).The expression levels of cyclin D1 protein in the untreated group,control siRNA group and Tiam1 siRNA group were 0.43 ± 0.02,0.41 ± 0.01 and 0.11 ± 0.02,respectively (P ＜ 0.05).The expression levels of p27 protein in the untreated group,control siRNA group and Tiam1 siRNA group were 0.10 ± 0.01,0.09 ± 0.02 and 0.20 ± 0.02,respectively (P ＜ 0.05).The ratios of early apoptotic cells in the untreated group,control siRNA group and Tiam1 siRNA group were (10 ± 0.9) ％,(10 ± 0.5) ％ and (27 ± 0.7) ％,respectively (P ＜ 0.01).The expression levels of Mcl-1 protein in EC9706 cells of untreated group,control siRNA group and Tiam1 siRNA group were 0.47 ±0.12,0.48 ±0.13 and 0.16 ±0.02,respectively (P ＜ 0.05).The expression levels of Bcl-2 protein in EC9706 cells of the untreated group,control siRNA group and Tiam1 siRNA group were 0.49 ±0.08,0.50 ±0.05 and 0.04 ± 0.03,respectively (P ＜ 0.05).The caspase-3 activities in the untreated group,comrol siRNA group and Tiam1 siRNA group were 2.3 ± 0.09,2.3 ± 0.10 and 16.0 ± 1.50,respectively; and that of caspase-9 were 2.3 ±0.08,2.3 ±0.11 and 14.5 ±0.9,respectively (P ＜0.05 for all).Conclusions Tiam1 siRNA can significantly inhibit the proliferation of esophageal cancer EC9706 cells,induce cell cycle arrest and cell apoptosis.These effects are related to the regulation of the expressions of cell cycle-related genes (cyclin D1 and p27) and cell apoptosis-related genes (Mcl-1,Bcl-1,caspase-3 and caspase-9) by Tiam1 siRNA.
