il18基因转染对小鼠卵巢癌ovhm体外增殖及体内成瘤的影响

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1、IL18基因转染对小鼠卵巢癌OVHM体外增殖及体内成瘤的影响作者：崔澂,郝淑维,李新伟,白小嘉,程建新,单保恩【摘要】　　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

2、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18mRNA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。15【关键

3、词】IL18基因转染OVHM增殖凋亡成瘤　　［Abstract］AIM:ToinvestigatetheeffectsofIL18genetransfectiononproliferation,apoptosisandtumorigenesisofmouseovariancancercelllineOVHM.METHODS:OVHMweretransfectedwithretrovirusencodingIL18gene(OVHM/IL18).ThewildOVHMandOVHMtransfectedwithvectorwithoutIL18gene(OVHM/LXSN)weree

4、nrolledasthecontrols.Theproliferation,cellcycleandapoptosisweremeasuredbyMTTandFCMrespectively.Thenudemiceweresubcutaneouslyinoculatedwiththethreedifferentcellsrespectively.Theobservationoftransplantedtumor’sgrowthandcalculationoftumorigenesisratiowasmade.TheexpressionofIL18mRNAintumortissueswasd

5、etectedbyRTPCR,andtheapoptosisoftumorcellintissuewasanalyzedbyelectronmicroscopeandFCM.RESULTS:Thethreecellsculturedinvitroshowednoapoptoticpeaks,andnodifferencesinproliferationandcellcycle(AllP>0.05).Followinginoculation,theratiosoftumorigenesisweresimilarinallthethreegroups.ButinOVHM/IL18gr

6、oup,thelatentperiodoftumorigenesiswaslongerwithslowertumorgrowthrateandpositiveexpressionofIL18mRNAintumortissue.Itwasalsoobservedthatinthetumorcellsfromnudemiceinoculated15byOVHM/IL18,thecellsinphaseofG0/G1increasedwithtypicalmorphologyofapoptosis,andthoseinSphasedecreasedwithdecreasedprolifera

7、tionindexandincreasedapoptosisindex(AllP<0.01).CONCLUSION:AlthoughIL18genehasnocytotoxiceffectsinvitro,itsinhibitingeffectsontumorigenesisofOVHMinvivowereexertedbyinterdictingcellcycleandacceleratingapoptosis