中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
11期
1700-1703
,共4页
食管癌%血管内皮生长因子%基因转染
食管癌%血管內皮生長因子%基因轉染
식관암%혈관내피생장인자%기인전염
Esophageal cancer%Vascular endothelial growth factor%Gene transfection
目的 观察血管内皮生长因子(VEGF)反义RNA对EC9706人食管癌细胞的抑制作用.方法 用脂质体法将反义VEGFcDNA质粒转染至EC9706人食管癌细胞,用噻唑蓝还原法(MTT法)检测EC9706细胞增殖,免疫组化SABC和逆转录-聚合酶链反应(RT-PCR)技术检测VEGF蛋白和VEGF mRNA表达水平.流式细胞术检测细胞凋亡和周期分布.并将转基因EC9706细胞接种于BALB/C裸鼠后肢皮下,4周后观察皮下成瘤情况.结果 被反义VEGFcDNA质粒转染的EC9706食管癌细胞有外源性VEGF反义基因的整合及表达,该细胞VEGF mRNA及蛋白的表达水平降低,但细胞生长增殖能力和增殖周期无明显改变,未发生明显凋亡现象;接种裸鼠28 d后,EC9706-wt组、EC9706-A组及EC9706-E组皮下移植瘤的潜伏期分别为(5.8±2.4)、(12.4±3.6)、(5.3±2.2)d,瘤体重量分别为(2.83 ±0.32)、(0.87±0.14)、(2.62 ±0.68)g,EC9706-A组与其他两组比较差异均有统计学意义(P＜0.05).结论 VEGF反义RNA可抑制EC9706食管癌细胞VEGF表达和裸鼠体内肿瘤生长.
目的 觀察血管內皮生長因子(VEGF)反義RNA對EC9706人食管癌細胞的抑製作用.方法 用脂質體法將反義VEGFcDNA質粒轉染至EC9706人食管癌細胞,用噻唑藍還原法(MTT法)檢測EC9706細胞增殖,免疫組化SABC和逆轉錄-聚閤酶鏈反應(RT-PCR)技術檢測VEGF蛋白和VEGF mRNA錶達水平.流式細胞術檢測細胞凋亡和週期分佈.併將轉基因EC9706細胞接種于BALB/C裸鼠後肢皮下,4週後觀察皮下成瘤情況.結果 被反義VEGFcDNA質粒轉染的EC9706食管癌細胞有外源性VEGF反義基因的整閤及錶達,該細胞VEGF mRNA及蛋白的錶達水平降低,但細胞生長增殖能力和增殖週期無明顯改變,未髮生明顯凋亡現象;接種裸鼠28 d後,EC9706-wt組、EC9706-A組及EC9706-E組皮下移植瘤的潛伏期分彆為(5.8±2.4)、(12.4±3.6)、(5.3±2.2)d,瘤體重量分彆為(2.83 ±0.32)、(0.87±0.14)、(2.62 ±0.68)g,EC9706-A組與其他兩組比較差異均有統計學意義(P＜0.05).結論 VEGF反義RNA可抑製EC9706食管癌細胞VEGF錶達和裸鼠體內腫瘤生長.
목적 관찰혈관내피생장인자(VEGF)반의RNA대EC9706인식관암세포적억제작용.방법 용지질체법장반의VEGFcDNA질립전염지EC9706인식관암세포,용새서람환원법(MTT법)검측EC9706세포증식,면역조화SABC화역전록-취합매련반응(RT-PCR)기술검측VEGF단백화VEGF mRNA표체수평.류식세포술검측세포조망화주기분포.병장전기인EC9706세포접충우BALB/C라서후지피하,4주후관찰피하성류정황.결과 피반의VEGFcDNA질립전염적EC9706식관암세포유외원성VEGF반의기인적정합급표체,해세포VEGF mRNA급단백적표체수평강저,단세포생장증식능력화증식주기무명현개변,미발생명현조망현상;접충라서28 d후,EC9706-wt조、EC9706-A조급EC9706-E조피하이식류적잠복기분별위(5.8±2.4)、(12.4±3.6)、(5.3±2.2)d,류체중량분별위(2.83 ±0.32)、(0.87±0.14)、(2.62 ±0.68)g,EC9706-A조여기타량조비교차이균유통계학의의(P＜0.05).결론 VEGF반의RNA가억제EC9706식관암세포VEGF표체화라서체내종류생장.
Objective To investigate the inhibitory effect of vascular endothelial growth factor (VEGF) antisense RNA on the growth of human esophageal cancer EC9706 cells in vitro. Methods The plasmid carrying VEGF antisense cDNA was transfected into EC9706 esophageal cancer cells, and the expression of VEGF was confirmed by reverse transcription-polymerase chain reaction (RT-PCR). The expression level of VEGF mRNA and protein was examined in antisense group by RT-PCR and immunohistochemistry staining, respectively. The cell growth rate was detected by MTT assay. Apotosis and cell cycle in transfected cells were measured by flow cytometry. Results The expression of exogenous antisense VEGF mRNA was confirmed in transfected cells, and the VEGF protein and endogenous VEGF mRNA were significantly decreased. The growth rate of transfected cells was not inhibited. Apototic cells were not found in transfected cells. Twenty-eight days after inoculation in nude mice in EC9706-wt group, EC9706-A group and EC9706-E group, latency of subcutaneous tumors was (5.8 ± 2. 4 ), ( 12. 4 ± 3.6) and (5.3 ± 2. 2) days, tumor weight was (2. 83 ± 0. 32 ), ( 0. 87 ± 0. 14 ) and ( 2. 62 ± 0. 68 ) g, respectively. There was significant difference between EC9706-A group and other groups ( P ＜ 0. 05). Conclusion VEGF antisense RNA could decrease the expression of VEGF in esophageal cancer cells and cell proliferation in vivo, which may provide a useful theory basis for gene therapy of esophageal cancer.
