中国实用医刊
中國實用醫刊
중국실용의간
CENTRAL PLAINS MEDICAL JOURNAL
2014年
20期
63-66
,共4页
TPX2%siRNA%细胞增殖%细胞周期%细胞凋亡
TPX2%siRNA%細胞增殖%細胞週期%細胞凋亡
TPX2%siRNA%세포증식%세포주기%세포조망
TPX2%siRNA%Cell proliferation%Cell cycle%Cell apoptosis
目的：研究 siRNA 沉默 TPX2对食管鳞癌 EC1细胞增殖、凋亡和侵袭的影响。方法构建针对TPX2的 siRNA 载体，利用 LipofectamineTM2000转染食管癌 EC1细胞。采用 RT-PCR 和 Western blot 分析 TPX2、Bcl-2、Cyclin D1的变化。用 MTT 法测定 TPX2 siRNA 对 EC1细胞增殖的影响，流式细胞术检测对 EC1细胞凋亡的影响，细胞体外侵袭实验检测对 EC1细胞侵袭、转移的影响。结果靶向 TPX2基因的 siRNA 的双链寡核苷酸片段克隆入 pRNAT-U6.1载体，经测序分析插入片段正确。RT-PCR 和 Western-blot 检测显示，转染 TPX2 siRNA 载体的细胞 TPX2、Bcl-2、Cyclin D1表达下调。TPX2 siRNA 能够抑制 EC1细胞的增殖，降低穿透 Matrigel 的肿瘤细胞数（P ﹤0.05），诱导肿瘤细胞凋亡（P ﹤0.01）。结论 siRNA 沉默食管癌 EC1细胞 TPX2基因表达能有效地诱导肿瘤细胞的凋亡，并抑制肿瘤细胞的增殖和侵袭、转移。
目的：研究 siRNA 沉默 TPX2對食管鱗癌 EC1細胞增殖、凋亡和侵襲的影響。方法構建針對TPX2的 siRNA 載體，利用 LipofectamineTM2000轉染食管癌 EC1細胞。採用 RT-PCR 和 Western blot 分析 TPX2、Bcl-2、Cyclin D1的變化。用 MTT 法測定 TPX2 siRNA 對 EC1細胞增殖的影響，流式細胞術檢測對 EC1細胞凋亡的影響，細胞體外侵襲實驗檢測對 EC1細胞侵襲、轉移的影響。結果靶嚮 TPX2基因的 siRNA 的雙鏈寡覈苷痠片段剋隆入 pRNAT-U6.1載體，經測序分析插入片段正確。RT-PCR 和 Western-blot 檢測顯示，轉染 TPX2 siRNA 載體的細胞 TPX2、Bcl-2、Cyclin D1錶達下調。TPX2 siRNA 能夠抑製 EC1細胞的增殖，降低穿透 Matrigel 的腫瘤細胞數（P ﹤0.05），誘導腫瘤細胞凋亡（P ﹤0.01）。結論 siRNA 沉默食管癌 EC1細胞 TPX2基因錶達能有效地誘導腫瘤細胞的凋亡，併抑製腫瘤細胞的增殖和侵襲、轉移。
목적：연구 siRNA 침묵 TPX2대식관린암 EC1세포증식、조망화침습적영향。방법구건침대TPX2적 siRNA 재체，이용 LipofectamineTM2000전염식관암 EC1세포。채용 RT-PCR 화 Western blot 분석 TPX2、Bcl-2、Cyclin D1적변화。용 MTT 법측정 TPX2 siRNA 대 EC1세포증식적영향，류식세포술검측대 EC1세포조망적영향，세포체외침습실험검측대 EC1세포침습、전이적영향。결과파향 TPX2기인적 siRNA 적쌍련과핵감산편단극륭입 pRNAT-U6.1재체，경측서분석삽입편단정학。RT-PCR 화 Western-blot 검측현시，전염 TPX2 siRNA 재체적세포 TPX2、Bcl-2、Cyclin D1표체하조。TPX2 siRNA 능구억제 EC1세포적증식，강저천투 Matrigel 적종류세포수（P ﹤0.05），유도종류세포조망（P ﹤0.01）。결론 siRNA 침묵식관암 EC1세포 TPX2기인표체능유효지유도종류세포적조망，병억제종류세포적증식화침습、전이。
Objective To study the effect of TPX2 siRNA on proliferation,apoptosis and inva-sion of esophageal carcinoma cell line EC1. Methods The siRNA vectors aiming to TPX2 gene were constructed. TPX2 siRNA was transfected into EC1 cells by LipofectamineTM2000. Changes of TPX2, Bcl-2 and Cyclin D1 were analyzed by Western blot and RT-PCR. Effect of TPX2 siRNA on EC1 cells proliferation was determined by MTT. Effect of TPX2 siRNA on EC1 cell apoptosis was detected by the flow cytometry. Boyden chamber was used to evaluate the invasion and metastasis capabilities of EC1 cells. Results The double strands oli-gonucleotide of siRNA aiming to TPX2 was successfully cloned into the pRNAT-U6. 1 vector,and the tar-get sequence is coincide with the design. RT-PCR and Western blotting detection demonstrate that the ex-pression level of TPX2,Bcl-2 and Cyclin D1 gene obviously decreased in EC1 cells transfected with TPX2 siRNA expression vectors,TPX2 siRNA could suppress the proliferation of EC1 cells. The average amount of cells penetrating Matrigel was significantly decreased(P ﹤ 0. 05). Silencing the TPX2 induced the apoptosis of esophageal carcinoma cell Line EC1(P ﹤ 0. 01). Conclusions TPX2 siRNA can induce the apoptosis of esophageal carcinoma cell line EC1. And it can suppress the proliferation,invasion and metastasis of EC1 cells.
