TNF相关凋亡诱导配体基因治疗小鼠肝细胞肿瘤

Effect of TNF-Related Apoptosis Inducing Ligand on Hepatocellular Carcinoma in Mice

目的:探讨经化疗药物作用后的H22细胞对TRAIL诱导凋亡的敏感性,观察TRAIL、纤黏连蛋白及内皮抑素的真核表达质粒pTRAIL,pCH510,pES联合化疗抑制小鼠肿瘤生长的作用。方法:pTRAIL转染BHK细胞后,加入分别经MMC,ADM,5-FU处理的H22细胞,MTT法检测pTRAIL对H22细胞的生长抑制作用,流式细胞仪分析H22细胞凋亡。在小鼠种植瘤体内注射MMC与pTRAIL,pCH510,pES,观察其对肿瘤生长的抑制作用。结果:pTRAIL能抑制H22生长并诱导细胞凋亡,与MMC,ADM及5-FU联合,H22的凋亡百分数分别为28.1%(P<0.01),22.5%(P<0.01)及47%(P>0.05)。体内,MMC+pTRAIL+pES+pCH510联合能有效抑制肝细胞肿瘤生长,肿瘤生成率为37.5%。结论:MMC或ADM作用后的残存H22肿瘤细胞对pTRAIL敏感,MMC+pTRAIL+pES+pCH510具有协同抑瘤效应,能将小鼠肝细胞肿瘤抑制在组织学水平。

Objective: To evaluate the sensibility of apoptosis to TRAIL on the residual H22 tumor cells after chemo-therapeutic agents treatment and the synergic therapeutic efficiency of pCH510, pTRAIL, and pES in combination with chemotherapeutic agent on mice tumor. Methods: Carrying full length of TRAIL gene, the eukaryotic expression plasmid of pTRAIL was transferred into BHK cells. The mouse hepatocellular carcinoma cell line of H22 which had been treated with ADM, MMC, or 5-FU were mixed with BHK cells. The inhibitory effect of pTRAIL in combination with chemotherapeutic agents was detected by MTT method. The percentage of apoptotic cells and cell cycle of residual H22 cells were analyzed by flow cytometry. The tumor model was made by inoculated with H22 hepatocarcinoma cells in mice. After injection of pCH510, pTRAIL, pES or MMC into intratumor, The therapeutic effects on tumor growth were assessed. Results: pTRAIL could inhibit the growth of H22 tumor cells and induce them to apoptosis. The percentage of apoptotic cells of TRAIL in combination with ADM, MMC, or 5-FU on H22 tumor cells was 28.1% (P <0.01) , 22.5% (P<0.01), and 47% (P >0.05). The tumor was effectively inhibited by MMC + pCH510 + pTRAIL + pES . And the percentage of tu-morogenesis was 37.5%. Residual tumor cells were scattered and mixed with immune cells from histological detection. Conclusion: Residual H22 tumor cells after ADM or MMC treatment is apt to be induced to apoptosis by pTRAIL. MMC + pCH510 + pTRAIL + pES is a great powerful modality for tumor synergic treatment.