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目的:分析转染IL-18基因后,对小鼠卵巢癌OVHM细胞体外增殖、凋亡及体内成瘤的影响,初步探讨IL-18基因转染治疗卵巢癌的应用价值。方法:将逆转录病毒携带的小鼠IL-18基因成功转染至OVHM(OVHM/IL-18),以空载体转染的OVHM(OVHM/LXSN)和野生型(OVHM)作为对照。分别采用MTT、FCM检测体外培养细胞的增殖、凋亡及周期分布。于裸鼠皮下分别接种细胞,观察各组种植瘤生长情况,计算成瘤率;RT-PCR检测瘤组织中IL-18mRNA表达;FCM和电镜分析肿瘤细胞增殖、凋亡状况。结果:3组细胞的体外增殖未见明显差异,均无明显凋亡现象,增殖指数、细胞周期分布均无明显差异(均P>0.05)。接种后,3组裸鼠的成瘤率相同,但OVHM/IL-18组成瘤时间较晚,随瘤龄增长肿瘤生长缓慢,瘤组织中IL-18 mRNA阳性表达。OVHM/IL-18组裸鼠种植瘤细胞可见典型的凋亡现象,G0/G1期细胞明显增多,S期细胞明显减少,增殖指数明显降低,凋亡指数明显增高(P<0.01)。结论:IL-18基因成功转染后,虽对体外卵巢癌细胞无直接毒性,但可能在体内借助非直接杀伤的其他途径,通过阻断细胞周期、促进肿瘤细胞凋亡等抑制体内成瘤。
OBJECTIVE: To analyze the effect of transfection of IL-18 gene on the proliferation, apoptosis and in vivo tumorigenesis of ovarian cancer OVHM cells in vitro and to investigate the value of IL-18 gene transfection in the treatment of ovarian cancer. Methods: Mouse IL-18 gene was transfected into OVHM (OVHM / IL-18) and OVHM (OVHM / LXSN) and wild type (OVHM) transfected with empty vector were used as controls. The proliferation, apoptosis and cycle distribution of cultured cells were detected by MTT and FCM respectively. The cells were inoculated subcutaneously in nude mice to observe the growth of implanted tumors in each group, and the tumor formation rate was calculated. The expression of IL-18 mRNA in tumor tissues was detected by RT-PCR, and the proliferation and apoptosis of tumor cells were analyzed by FCM and electron microscope. Results: There was no significant difference in proliferation between the 3 groups of cells in vitro. There was no significant apoptosis in the 3 groups. There was no significant difference in proliferation index and cell cycle between the 3 groups (all P> 0.05). After inoculation, the nude mice had the same rate of tumorigenesis, but the OVHM / IL-18 group formed later, the tumor growth was slow with tumor growth, and the expression of IL-18 mRNA was positive in the tumor tissue. The typical apoptotic cells were observed in OVHM / IL-18 group. The number of cells in G0 / G1 phase was significantly increased, the number of S phase cells was significantly decreased, the proliferation index was significantly decreased and the apoptosis index was significantly increased (P <0.01). CONCLUSION: Although IL-18 gene is not directly toxic to ovarian cancer cells in vitro, IL-18 gene may inhibit the tumorigenesis in vivo by blocking the cell cycle, promoting apoptosis of tumor cells and so on in other ways by indirect killing.