中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
32期
2292-2295
,共4页
张靖%付彦超%康春生%张庆瑜%王涛%张洁
張靖%付彥超%康春生%張慶瑜%王濤%張潔
장정%부언초%강춘생%장경유%왕도%장길
蛋白激酶类%环氧化酶2%胃肿瘤%腺病毒载体
蛋白激酶類%環氧化酶2%胃腫瘤%腺病毒載體
단백격매류%배양화매2%위종류%선병독재체
Protein kinases%Cyclooxygenase 2%Stomach neoplasms%Adenovirus vector
目的 研究靶向蛋白激酶B1(PKB1/Akt1)和环氧合酶2(COX-2)短发夹RNA(shRNA)腺病毒载体对SGC-7901人胃腺癌细胞生长的作用.方法 构建腺病毒载体pGSadeno-Akt1+COX-2(rAd5-A+C),体外转染人胃癌细胞株SGC-7901后,噻唑蓝比色分析法(MTT法)和流式细胞术评价转染后胃癌细胞的增殖活性.裸鼠皮下荷瘤模型进一步观察rAd5-A+C对SGC-7901生长的抑制效果,并应用原位末端标记技术检测肿瘤细胞的原位凋亡.结果构建的rAd5-A+C重组腺病毒载体转染SGC-7901后,胃癌细胞的增殖活性明显被抑制.细胞周期分析显示对照组和空载组在G0/G1期,S期和G2/M期的细胞数占细胞总数分别为49.8%、35.2%、15.0%和50.8%、36.5%、12.7%;而治疗组在G0/G1期、s期和G2/M期的细胞数占细胞总数的68.1%、21.8%和10.1%.裸鼠皮下荷瘤模型显示rAd5-A+C可抑制肿瘤的生长和诱导细胞的凋亡.结论 腺病毒介导的Aktl和COX-2 shRNA可抑制人胃癌细胞的生长,这可能为胃癌靶向性联合基因治疗提供了新的策略.
目的 研究靶嚮蛋白激酶B1(PKB1/Akt1)和環氧閤酶2(COX-2)短髮夾RNA(shRNA)腺病毒載體對SGC-7901人胃腺癌細胞生長的作用.方法 構建腺病毒載體pGSadeno-Akt1+COX-2(rAd5-A+C),體外轉染人胃癌細胞株SGC-7901後,噻唑藍比色分析法(MTT法)和流式細胞術評價轉染後胃癌細胞的增殖活性.裸鼠皮下荷瘤模型進一步觀察rAd5-A+C對SGC-7901生長的抑製效果,併應用原位末耑標記技術檢測腫瘤細胞的原位凋亡.結果構建的rAd5-A+C重組腺病毒載體轉染SGC-7901後,胃癌細胞的增殖活性明顯被抑製.細胞週期分析顯示對照組和空載組在G0/G1期,S期和G2/M期的細胞數佔細胞總數分彆為49.8%、35.2%、15.0%和50.8%、36.5%、12.7%;而治療組在G0/G1期、s期和G2/M期的細胞數佔細胞總數的68.1%、21.8%和10.1%.裸鼠皮下荷瘤模型顯示rAd5-A+C可抑製腫瘤的生長和誘導細胞的凋亡.結論 腺病毒介導的Aktl和COX-2 shRNA可抑製人胃癌細胞的生長,這可能為胃癌靶嚮性聯閤基因治療提供瞭新的策略.
목적 연구파향단백격매B1(PKB1/Akt1)화배양합매2(COX-2)단발협RNA(shRNA)선병독재체대SGC-7901인위선암세포생장적작용.방법 구건선병독재체pGSadeno-Akt1+COX-2(rAd5-A+C),체외전염인위암세포주SGC-7901후,새서람비색분석법(MTT법)화류식세포술평개전염후위암세포적증식활성.라서피하하류모형진일보관찰rAd5-A+C대SGC-7901생장적억제효과,병응용원위말단표기기술검측종류세포적원위조망.결과구건적rAd5-A+C중조선병독재체전염SGC-7901후,위암세포적증식활성명현피억제.세포주기분석현시대조조화공재조재G0/G1기,S기화G2/M기적세포수점세포총수분별위49.8%、35.2%、15.0%화50.8%、36.5%、12.7%;이치료조재G0/G1기、s기화G2/M기적세포수점세포총수적68.1%、21.8%화10.1%.라서피하하류모형현시rAd5-A+C가억제종류적생장화유도세포적조망.결론 선병독개도적Aktl화COX-2 shRNA가억제인위암세포적생장,저가능위위암파향성연합기인치료제공료신적책략.
Objective To construct a short hairpin RNA (shRNA) adenovirus vector targeting Akt1 (protein kinase BI, PKBI/Akt1 ) and cyclooxygenase-2 (COX-2) and study its effects on the growth of SGC-7901 human gastric adenocarcinoma cell. Methods Adenovirus pGSadeno-Aktl + COX-2 (rAdS-A + C) vector was constructed and transfected into SGC-7901 cell. The proliferative activity of tumor cell was evaluated by MTT assay and flow cytometry in vitro, rAd5-HK and rAd5-A + C were injected into the established subcutancous SGC-7901 gastric adenocarcinoma in nude mice. During the observation period of 21 days, tumor volume was measured every 3 days to further observe the anti-tumor effects of rAd5-A + C on SGC-7901 cell and cell situ apoptosis was detected by TUNEL assay. Results After transfection of constructed adenovirus vector rAdS-A + C into SGC-7901 cell, cell proliferative activity in rAdS-A + C treatment group was significantly suppressed, and cell cycle indicated that control group SGC-7901 and no-load group rAd5-A + C cells in G0/G1 ,S and G2/M phases accounted for the total number of cells 49. 8%, 35.2%, 15.0% and 50. 8%, 36. 5%, 12. 7% respectively. While the treatment group rAdS-A +C in G0/ G1 ,S and G2/M phases accounted for the total number of cells 68. 1%, 21.8% and 10. 1% respectively. The tumor volume in treatment group was lower than that of control and no-load groups and the difference had statistical significance (F= 3.679, P =0. 043 ) and rAdS-A + C could induce the apoptosis of tumor cell. Conclusion Adenovirus-mediated Aktl and COX-2 shRNA can inhibit the growth of SGC-7901 human gastric adenocarcinoma cell. It may provide a new strategy for gastric cancer gene therapy.
