腹腔热化疗对大鼠卵巢癌细胞凋亡的影响

目的 观察顺铂腹腔热化疗对卵巢癌大鼠的疗效,并且从细胞凋亡角度进一步探讨腹腔热化疗治疗卵巢癌的机制,为临床应用腹腔热化疗治疗卵巢癌提供实验依据.方法 体外培养近交系Fischer 344大鼠低分化上皮性卵巢癌细胞株NuTu-19,将一定数昔的培养细胞分别注入24只雌性近交系Fischer344大鼠腹腔中,建立卵巢癌腹腔转移瘤动物模型;实验动物随机分组后,应用顺铂及不同加热方式进行治疗;治疗结束后处死大鼠,提取腹腔积液中游离卵巢癌细胞,应用流式细胞仪、PI染色检测其细胞周期比率及细胞凋亡率;取卵巢癌腹腔转移瘤组织进行免疫组化分析,检测凋亡相关基冈bcl-2及bax的表达.结果 ①所有实验动物均表现出不同程度的血性腹腔积液及腹腔转移瘤微小结节病灶;②顺铂或加热治疗均町提高大鼠腹腔积液中卵巢癌细胞的凋亡率（P〈0.05）,顺铂与微波持续加热同时应用后细胞凋亡率进一步升高（P〈0.05）;③顺铂或一次性加热治疗可降低大鼠腹腔积液中游离卵巢癌细胞G0/G1期比率（P〈0.05）;微波持续加热治疗或与顺铂同时应用均可降低腹腔积液中卵巢癌细胞S期细胞比率（P〈0.05）;④顺铂或微波持续加热治疗的大鼠腹腔转移瘤组织中bcl-2基因表达降低（P〈0.05）,bax基因表达升高（P〈0.05）,两种疗法联合治疗具有正向协同作用（P〈0.05）.结论 ①顺铂及不同加热方式治疗可通过促进细胞凋亡或抑制细胞进入增殖期从而起到治疗卵巢癌的作用,且顺铂联合微波持续加热治疗效果最佳.②顺铂及微波持续加热治疗均可降低卵巢癌组织中bcl-2基因表达,增加bax基因表达,从而诱导卵巢癌细胞凋亡,并且两法同时应用效果更佳.

Effect of intraperitoneal hyperthermic chemotherapy on apoptosis of rat ovarian cancer cells

Objective To observe the therapeutic efficacy for the application of cisplatin associating with microwave persistent heating to the rats with ovarian cancer in abdominal cavity metastasis, approach the further mechanisms of in- traperitoneal hyperthermic chemotherapy（IPHC） from the view of apoptosis and provide reference for clinical application of IPHC. Method Poorly differentiated epithelial cell line NuTu - 19 of Fischer 344 rat ovarian cancer was cultured in vitro. Then, we established ovarian cancer metastasis models by abdominal cavity injection in inbred line Fischer 344 rats. Cisplatin and different heat - treatment were carried out to heal the rats of randomization. After the therapy, all the rats were killed and the free tumor ceils in hydroperitoneum as well as the metastatic tumor tissues were taken. Histo- chemistry essay was used to detect the expressions of both bcl - 2 and bax genes in metastatic tumor tissues. While, anal- ysis of apoptosis rate and ratio of different cell cycle phases of the free tumor cells in hydroperitoneum were taken by the use of flow cytometry and PI staining method. Results （1）All the animals were displayed inequality hemorrhagic ascites and metastatic tumor focus of infection in abdominal cavity; （2）Cisplatin or microwave persistente heating treatment increased the apoptosis rate of the free tumor cells in hydroperitoneum （P 〈 0.05 ）, also, there was a positive synergistic effect of these two factors （P 〈 0.05 ） ; （3）The application of cisplatin or once heating therapy depressed the ratio of Go/ Gl - phase of the free tumor cells in hydroperitoneum （P 〈 0.05 ）. Microwave persistent heating treatment or therapeutic alliance of these two factors depressed the ratio of S - phase cells （P 〈 0.05 ） ; （4）The application of cisplatin or microwave persistent heating downregulated the expression of bcl - 2 gene （P 〈 0.05 ） and increased bax expression levels （ P〈 0.05 ） significantly. Moreover, therapeutic alliance of these two factors possessed a positive synergistic effect. Conclusion （1）Cisplatin or heating treatment elevate apoptosis rate of the free tumor cells in hydroperitoneum and cisplatin associating with microwave persistente heating make a further increasing in the apoptosis rate. （2）Cisplatin or microwave persistent heating and the two factors synergistic effects displaiy depressing expression of bcl - 2 gene and increasing bax gene, suggest inducing tumor cell apoptosis.