金雀异黄素联合阿霉素对卵巢癌A2780细胞增殖和荷瘤裸鼠肿瘤生长的影响

Study on the effect of genistein and adriamycin on the proliferation of human ovarian cell A2780 and growth of tumor bearing nude mice

目的探讨不同浓度的金雀异黄素联合阿霉素对卵巢癌A2780细胞增殖和荷瘤裸鼠肿瘤生长的影响。方法将卵巢癌A2780细胞制成细胞悬液后接种于裸鼠皮下,制备卵巢癌裸鼠模型。将40只造模成功的裸鼠随机分为4组,即生理盐水组、金雀异黄素联合阿霉素低剂量组、中剂量组、高剂量组,腹腔注射给药,每周1次。用药4周后,处死裸鼠,剥取肿瘤组织,称重,计算各组肿瘤抑制率。用免疫组化方法检测各组肿瘤组织中c-mye的表达。分别采用低、中、高剂量金雀异黄素联合阿霉素共同作用于卵巢癌A2780细胞,Western blot方法检测各组细胞内c-myc的表达,MTT检测各组细胞的增殖能力,细胞黏附实验与transwell法检测各组细胞的侵袭能力,流武细胞术检测各组细胞周期及凋亡率。结果与生理盐水组相比,金雀异黄素联合阿霉素中、高剂量组的抑瘤率分别为(40.26±4.25)%、(44.81±5.74)%,差异均有统计学意义(P<0.05)。免疫组化结果显示,金雀异黄素联合阿霉素中、高剂量组能明显抑制c-myc的活化。金雀异黄素联合阿霉素中、高剂量组能显著降低卵巢癌A2780细胞中c-myc的表达,抑制细胞增殖及降低侵袭能力,显著升高G0/G1期细胞(P<0.05),降低S期细胞(P<0.05),且细胞凋亡率显著高于对照组(P<0.05)。结论一定剂量的金雀异黄素联合阿霉素可在体内抑制卵巢癌裸鼠移植瘤的生长及体外抑制卵巢癌A2780细胞的增殖、降低侵袭能力及促进细胞凋亡,其机制可能是通过降低c-myc蛋白的表达,影响c-myc参与调控的癌细胞增殖和转移过程。

Objective To study the effects of genistein and adriamycin on the proliferation of human ovarian cell A2780 and growth of tumor bearing nude mice.Methods The ovarian cancer cell A2780 suspension were inoculated into nude mice (n=40 )to establish the animal models with ovarian tumor.These model rats were randomly divided into four groups,saline group and different genistein concentrations combined with adriamycin treatment group (high-dose,middle-dose and low-dose group)by intraperitoneal injection once a week.After treatment for four weeks,the ovarian tumors in all rats were peeled off and weighted for the measurement of tumor inhibition rate.The expression of c-myc in ovarian tumors were detected by immunohistochemistry.In vitro,after affected by different genistein concentrations combined with adriamycin,the expression of c-myc in A2780 cell of each group were detected by western blot.The proliferation of A2780 cell were determined by MTT method,and the invasiveness of cell by cell adhesion and transwell assay,the apoptosis rate and cell cycle were measured by flow cytometry. Results The tumor inhibition rates in middle-dose (40.26 ± 4.25)% and high-dose groups[,(44.81 ±5.74)%])were significantly lower than saline group (P<0.05).Immunohistochemistry results showed that c-myc monoclonal antibody in middle-dose and high-dose groups can significantly inhibit the activation of c-myc in ovarian carcinoma tissue.Compared with control group,the expression of c-myc in A2780 cell were significantly decreased,the proliferation and invasiveness of cell were inhibited,cells in G0/G1 phase were significantly increased(P<0.05)and in S phase were(P<0.05)decreased,and the apoptosis were induced by genistein combined with adriamycin in middle-dose and high-dose (P <0.05 ).Conclusion Genistein combined with adriamycin in certain doses could inhibit the proliferation and invasiveness of ovarian carcinoma,and induce its apoptosis.The mechanism may be related to the decreased expression of c-myc.