海南医学
海南醫學
해남의학
HAINAN MEDICAL JOURNAL
2014年
13期
1880-1883
,共4页
曹佳%贺思佳%徐雷鸣%李兆申
曹佳%賀思佳%徐雷鳴%李兆申
조가%하사가%서뢰명%리조신
TM4SF1%人脐静脉内皮细胞%管腔形成
TM4SF1%人臍靜脈內皮細胞%管腔形成
TM4SF1%인제정맥내피세포%관강형성
TM4SF1%HUVECs%Tube formation
目的：研究TM4SF1对内皮细胞体外管腔形成的影响。方法应用qRT-PCR方法检测人脐静脉内皮细胞(HUVECs)中TM4SF1的mRNA表达量；应用siRNA方法瞬时转染HUVECs，采用qRT-PCR方法检测转染48 h与72 h后TM4SF1的沉默效果，选择最佳的转染时间，应用In Vitro Angiogenesis Assay Kit观察siControl与siTM4SF1转染后HUVECs体外管腔形成情况。结果 qRT-PCR实验结果显示，HUVECs中表达TM4SF1的RQ值为(0.71±0.11)，已知高表达TM4SF1胰腺癌细胞株MPanc96表达TM4SF1的RQ值为(0.56±0.13)，两种细胞中TM4SF1表达量差异无统计学意义(P>0.05)；siRNA瞬时转染HUVECs 48 h与72 h后，TM4SF1表达分别下降了47.06%与93.14%，HUVECs瞬时转染72 h后进行体外管腔形成实验，通过对管腔形成情况进行评级，siControl转染后HUVECs管腔形成情况为5级(多个闭合管状结构相连形成筛状)，siTM4SF1转染后为2级(细胞排列形成夹角)。结论 TM4SF1在HUVECs中高表达，沉默TM4SF1后可以明显抑制HUVECs的体外管腔形成，提示TM4SF1与肿瘤血管形成相关。
目的：研究TM4SF1對內皮細胞體外管腔形成的影響。方法應用qRT-PCR方法檢測人臍靜脈內皮細胞(HUVECs)中TM4SF1的mRNA錶達量；應用siRNA方法瞬時轉染HUVECs，採用qRT-PCR方法檢測轉染48 h與72 h後TM4SF1的沉默效果，選擇最佳的轉染時間，應用In Vitro Angiogenesis Assay Kit觀察siControl與siTM4SF1轉染後HUVECs體外管腔形成情況。結果 qRT-PCR實驗結果顯示，HUVECs中錶達TM4SF1的RQ值為(0.71±0.11)，已知高錶達TM4SF1胰腺癌細胞株MPanc96錶達TM4SF1的RQ值為(0.56±0.13)，兩種細胞中TM4SF1錶達量差異無統計學意義(P>0.05)；siRNA瞬時轉染HUVECs 48 h與72 h後，TM4SF1錶達分彆下降瞭47.06%與93.14%，HUVECs瞬時轉染72 h後進行體外管腔形成實驗，通過對管腔形成情況進行評級，siControl轉染後HUVECs管腔形成情況為5級(多箇閉閤管狀結構相連形成篩狀)，siTM4SF1轉染後為2級(細胞排列形成夾角)。結論 TM4SF1在HUVECs中高錶達，沉默TM4SF1後可以明顯抑製HUVECs的體外管腔形成，提示TM4SF1與腫瘤血管形成相關。
목적：연구TM4SF1대내피세포체외관강형성적영향。방법응용qRT-PCR방법검측인제정맥내피세포(HUVECs)중TM4SF1적mRNA표체량；응용siRNA방법순시전염HUVECs，채용qRT-PCR방법검측전염48 h여72 h후TM4SF1적침묵효과，선택최가적전염시간，응용In Vitro Angiogenesis Assay Kit관찰siControl여siTM4SF1전염후HUVECs체외관강형성정황。결과 qRT-PCR실험결과현시，HUVECs중표체TM4SF1적RQ치위(0.71±0.11)，이지고표체TM4SF1이선암세포주MPanc96표체TM4SF1적RQ치위(0.56±0.13)，량충세포중TM4SF1표체량차이무통계학의의(P>0.05)；siRNA순시전염HUVECs 48 h여72 h후，TM4SF1표체분별하강료47.06%여93.14%，HUVECs순시전염72 h후진행체외관강형성실험，통과대관강형성정황진행평급，siControl전염후HUVECs관강형성정황위5급(다개폐합관상결구상련형성사상)，siTM4SF1전염후위2급(세포배렬형성협각)。결론 TM4SF1재HUVECs중고표체，침묵TM4SF1후가이명현억제HUVECs적체외관강형성，제시TM4SF1여종류혈관형성상관。
Objective To investigate the effect of TM4SF1 on capillary-like tube formation in human umbili-cal vein endothelial cells (HUVECs) in vitro. Methods The mRNA expression of TM4SF1 in HUVECs was detected by quantitative reverse transcription-PCR (qRT-PCR). HUVECs were transiently transfected with siRNA, and the si-lence of TM4SF1 was determined by qRT-PCR at 48 h or 72 h after transfection to confirm the time of the best silenc-ing effect. The capillary-like tube formation was detected by In Vitro Angiogenesis Assay Kit and compared between siControl and siTM4SF1. Results The results of qRT-PCR showed the expression of TM4SF1 in HUVECs and in MPanc96, a pancreatic cancer cell line, as well. There were no significant difference in RQ values between HUVECs and MPanc96 by Mann-Whitney U test [(0.71±0.11)vs (0.56±0.13), P>0.05]. The expressions of TM4SF1 in HUVECs decreased by 47.06%and 93.14%at 48 h and 72 h after transiently transfection. The capillary-like tube formation was detected in HUVECs at 72 h after transiently transfection by In Vitro Angiogenesis Assay Kit, and scored for the de-gree subsequently. The tube formation of HUVECs transfected by siControl was grade 5 (Closed polygons begin to form, and complex mesh like structures develop), while it was grade 2 when it was transfected by siTM4SF1 (Some cells formed angles but no sprouting). Conclusion TM4SF1 was highly expressed in HUVECs. The capillary-like tube formation of HUVECs significantly decreased after silencing of TM4SF1 in vitro. This study indicated that TM4SF1 might be involved in angiogenesis in cancer.
