增免抑瘤方联合顺铂对铂类耐药卵巢癌抑瘤率影响的实验研究

目的观察增免抑瘤方对顺铂耐药卵巢癌裸鼠皮下移植瘤生长的抑制作用,并探讨其可能的作用机制。方法采用顺铂耐药人卵巢癌细胞株COC1/DDP构建裸鼠皮下移植瘤模型,随机分为中药高、中、低剂量组,顺铂组,联合组(顺铂联合中药),对照组(生理盐水),连续给药3周,剥取瘤体计算瘤重及抑瘤率;定量RT-PCR及免疫组化测定Bcl相关X连锁蛋白(Bcl-associated x protein,Bax)、B细胞淋巴瘤/白血病基因-2(B-cell lymphoma/leukemia-2,Bcl-2)的表达;电镜观察瘤体细胞超微结构。结果联合组抑瘤率最高,为(59.26±6.86)%,与其他各组比较差异均有统计学意义(P<0.01)。RT-PCR结果显示:促凋亡基因Bax mRNA联合组表达最高,对照组表达最低(P<0.01)。抑凋亡基因Bcl-2mRNA对照组表达最高,中药高剂量组表达最低,联合组Bcl-2mRNA的表达低于顺铂组(P<0.01)。免疫组化结果显示:联合组Bax蛋白的表达最高,Bcl-2蛋白的表达最低,与其他各组比较差异均有统计学意义(P<0.01)。电镜下中药各剂量组与联合组均可见凋亡中、晚期的表现。结论增免抑瘤方辅助化疗可逆转耐药卵巢癌裸鼠对顺铂的耐药性,增强其抑瘤作用。其机制与上调Bax的表达,下调Bcl-2的表达,促进耐药卵巢癌细胞的凋亡有关。

Effects of Zengmian Yiliu Recipe Combined Cisplatin on the Tumor Inhibition Rate in Platinum-resistant Ovarian Cancer

Objective To observe the effects of Zengmian Yiliu Recipe(ZYR) combined cisplatin on the growth of subcutaneous tumor in nude mice with platinum-resistant ovarian cancer,and to explore its possible mechanisms.Methods The model of ovarian cancer subcutaneous tumor was established in nude mice using platinum-resistant ovarian cancer line COC1/DDP.The mice were randomly divided into six groups,i.e.,the high Chinese materia medica(CMM) group,the medium CMM group,the low CMM group,the cisplatin group(DDP),the combined treatment group(with DDP combined CMM),and the control group(with normal saline).The medication lasted for 3 successive weeks.The tumor weight and the tumor inhibition rate were calculated.The expressions of Bcl-associated x protein(Bax) and B-cell lymphoma/leukemia-2(Bcl-2) were detected using quantitative RT-PCR and immunohistochemical assay.The ultra-structure of tumor cells was observed by electron microscopy.Results The tumor inhibition rate was the highest in the combined treatment group,being(59.26±6.86)%,showing statistical difference when compared with the rest groups(P<0.01).Results of RT-PCR showed the Bax mRNA expression was the highest in the combined treatment group and the lowest in the control group(P<0.01).Anti-apoptotic gene Bcl-2 mRNA expression was the highest in the control group and the lowest in the high CMM group(P<0.01).The Bcl-2 mRNA expression was lower in the combined treatment group than in the cisplatin group(P<0.01).Immunohistochemical results showed the Bax protein expression was the highest and the expression of Bcl-2 was the lowest in the combined treatment group,showing statistical difference when compared with the rest groups(P<0.01).The middle-and late-stage manifestations of apoptosis could be seen in each CMM group and the combined treatment group under electron microscope.Conclusions ZYR combined with chemotherapy could reverse the cisplatin-resistance of resistant ovarian cancer nude mice,and enhance its tumor inhibitory effect.Its mechanisms were correlated with up-regulating the expression of Bax,down-regulating the expression of Bcl-2,and promoting cisplatin resistant ovarian cancer cell apoptosis.