CAJ | 학술논문

目的观察苦参碱对TIM2基因修饰小鼠H22肝癌细胞瘤苗的体内作用.方法以携带小鼠TIM2基因的真核表达载体脂质体法转染H22细胞,经体外稳定筛选获得TIM2转基因H22细胞瘤苗.建立小鼠肝癌移植瘤模型,接种TIM2转基因H22细胞瘤苗(H22-TIM2组),观察成瘤情况,并设空载体转染的H22细胞稳定表达株和H22细胞作为对照(H22-EGFP组和H22组);同时,分别加用药物苦参碱治疗,观察苦参碱对不同处理组小鼠肿瘤生长情况的影响.结果成功得到TIM2转基因修饰H22细胞,鉴定有TIM2基因mRNA及EGFP的稳定表达.免疫接种小鼠后,在小鼠体内的成瘤率为41.6%,明显低于H22组和H22-EGFP组(后两者成瘤率在91.6%以上)(P＜0.01),小鼠肿瘤的生长速率也较其他各组明显缓慢,实验结束时H22-TIM2组小鼠肿瘤平均体积为(31.34±9.21)mm3,明显小于H22.EGFP组和H22组[(98.25±25.23)mm3和(114.08±36.45)mm3;P＜0.01].接种H22-TIM2细胞对小鼠肿瘤的抑制率为69.2%,加用苦参碱后,抑瘤率达90.6%,明显高于单用苦参碱组(67.5%)及H22-EGFP和苦参碱联用组(70.8%).结论 TIM2基因修饰H22细胞可显著降低H22肝癌细胞在小鼠体内的致瘤性,抑制小鼠体内H22移植瘤的发生和发展,而苦参碱可明显改善其体内抗癌活性.
목적 관찰고삼감대TIM2기인수식소서H22간암세포류묘적체내작용.방법 이휴대소서TIM2기인적진핵표체재체지질체법전염H22세포,경체외은정사선획득TIM2전기인H22세포류묘.건립소서간암이식류모형,접충TIM2전기인H22세포류묘(H22-TIM2조),관찰성류정황,병설공재체전염적H22세포은정표체주화H22세포작위대조(H22-EGFP조화H22조);동시,분별가용약물고삼감치료,관찰고삼감대불동처리조소서종류생장정황적영향.결과성공득도TIM2전기인수식H22세포,감정유TIM2기인mRNA급EGFP적은정표체.면역접충소서후,재소서체내적성류솔위41.6%,명현저우H22조화H22-EGFP조(후량자성류솔재91.6%이상)(P＜0.01),소서종류적생장속솔야교기타각조명현완만,실험결속시H22-TIM2조소서종류평균체적위(31.34±9.21)mm3,명현소우H22.EGFP조화H22조[(98.25±25.23)mm3화(114.08±36.45)mm3;P＜0.01].접충H22-TIM2세포대소서종류적억제솔위69.2%,가용고삼감후,억류솔체90.6%,명현고우단용고삼감조(67.5%)급H22-EGFP화고삼감련용조(70.8%).결론 TIM2기인수식H22세포가현저강저H22간암세포재소서체내적치류성,억제소서체내H22이식류적발생화발전,이고삼감가명현개선기체내항암활성.
Objective To investigate the effects of matrine on the anti-tumor efficiency of TIM2gene-modified murine hepatocarcinoma H22 cells.Methods A combined eukaryotic expression vector DIRES2-EGFP-TIM2 was constructed and transfected into H22 cells by lipofectamin.The monoclone of positive H22-TIM2 cells and negative control H22-EGFP cells transfected with plRES2-EGFP vector were selected by G418 pressure and limited dilution method in turn and were inoculated to establish the tumorbearing mouse model.Next.matrine was administered to the tumor-bearing mice and the inhibitory effect of matrine was determined.Results The co-expression of EGFP protein and TIM2 gene was detected in H22 cells selected after TIM2 gene transfecion.After subcutaneous injection of H22-TIM2 cells,the rate of tumor formation(41%)was lower than that of H22 cells and H22-EGFP cells iniection(92%)in mice.The tumor growth was significantly inhibited in mice vaccinated with H22-TIM2 cells.After the experiment was completed,the volume of tumors in mice of H22-TIM2 group was 3 1.34±9.21 mm3,smaller than those in H22-EGFP group(98.25±25.23)mm3and H22 cells group(114.08±36.45)mm3(P＜0.01).Matrine dramatically enhanced the anti-tumor efficiency of TIM2 gene-modified H22 cells,with the highest tumor inhibitory rate(IR)90.6%among the H22-TIM2 group,matrine treatment group and H22-EGFP cells combined with matrine treatment group(69.2%,67.5%and 70.8%,respectively)in the experimental mice.Condusion The tumorigenesity of H22 cells has been markedly impaired after modification by TIM2 gene.Matrine can enhance its inhibitory effect on tumors of H22-TIM2 cells in vivo.These data indicate importance to further study on the biological role of TIM2 gene in tumor immunity and explore the molecular mechanism of matrine in suppressing of tumor growth.