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目的探讨溴隐亭对肝癌多药耐药逆转的作用机制。方法体外实验分亲本细胞HepG2组(A组),耐药细胞HepG2／A13M组(B组)和B组加溴隐亭称为C组。分别检测各组细胞内荧光强度变化,细胞P-糖蛋白、蛋白激酶C α蛋白表达情况,及5种肿瘤药物半数抑制浓度及耐药指数变化。分别将肝癌细胞:HepG2和耐药细胞HepG2／ADM原位种植入裸鼠肝脏内,称A组鼠和B组鼠,另设B组种植鼠给予溴隐亭灌冒治疗称C组鼠。B超观察种植瘤的生长情况,肿瘤大小为1．0cm左右时经腹腔化疗,2周后处死裸鼠计算肿瘤的体积和质量抑制率,观察种植瘤组织多药耐药基因1不敷出 mRNA表达情况,及治疗后肝癌细胞的凋亡指数。单独检测24只种植耐药细胞HepG2／ADM的裸鼠在服用溴隐亭前后分别注射的99mTc-MIBI在肝脏肿瘤部位的潴留情况。结果体外实验结果显示在溴隐亭浓度为10 μmol／L时罗丹明123 在细胞内的潴留率均明显增加,且呈时间依赖性,对阿霉素的耐药逆转以溴隐亭浓度为10μmol／L时最显著,逆转率为 82．6％。蛋白激酶C-α蛋白表达,C组与B组比较差异有统计学意义(q=5．37,P<0．01);但两组P-糖蛋白表达差异无统计学意义(q=1.86,P>0．05)。体内试验结果显示种植瘤的体积和质量抑制率比较,C组鼠明显高于B组鼠(q1= 5．89,q2=4．92,P<0．01),接近于A组鼠(q1=2．47,q2=3．02,P>0．05)。C组鼠与B组鼠多药耐药基因1 mRNA 表达差异无统计学意义(q=3．71,P>0．05)。化疗后肿瘤细胞凋亡率比较,C组鼠明显高于B组鼠(q=3．72, P<0．01)。口服溴隐亭后肝肿瘤组织对99mTc-MIBI的潴留指数明显提高(t=3．58,P<0．01)。结论溴隐亭通过抑制 P-糖蛋白的功能可有效的逆转肝癌多药耐药性。
Objective To investigate the mechanism of bromocriptine on the reversal of multidrug resistance in liver cancer. Methods The in vitro experiments were divided into HepG2 group (group A), HepG2 / A13M group (group B) and B group plus bromocriptine (group C). The change of intracellular fluorescence intensity, the expression of P-glycoprotein and protein kinase Cα, the half inhibitory concentration and the drug resistance index of five kinds of oncology were detected. HepG2 cells and HepG2 / ADM cells were inoculated into the liver of nude mice in situ, respectively. The rats in group A and group B were treated with bromocriptine. B ultrasound observation of the growth of implanted tumor, the tumor size of about 1.0cm by intraperitoneal chemotherapy, two weeks after the nude mice were sacrificed to calculate the size of the tumor and mass inhibition rate of implanted tumor tissue multi-drug resistance gene 1 makes up mRNA Expression, and apoptosis index of hepatoma cells after treatment. The retention of 99mTc-MIBI in 24 tumor-bearing HepG2 / ADM nude mice before and after taking bromocriptine was detected separately in the liver tumor site. Results The results of in vitro experiments showed that the intracellular retention rate of rhodamine 123 in bromocriptine concentration of 10 μmol / L was significantly increased in a time-dependent manner, and the resistance to doxorubicin was reversed with bromocriptine concentration of 10 μmol / L, the most significant reversal rate was 82.6%. Protein kinase C-αprotein expression, there was a significant difference between group C and group B (q = 5.37, P <0.01). However, there was no significant difference in the expression of P-glycoprotein between the two groups (q = 1.86 , P> 0.05). The results of in vivo experiments showed that the volume and mass inhibition rate of implanted tumors were significantly higher in group C than in group B (q1 = 5.89, q2 = 4.92, P <0.01) = 2.47, q2 = 3.02, P> 0.05). There was no significant difference in multidrug resistance 1 mRNA expression between group C and group B (q = 3.71, P> 0.05). The apoptosis rate of tumor cells after chemotherapy was significantly higher in group C than in group B (q = 3.72, P <0.01). The retention index of 99mTc-MIBI in oral tumor of bromocriptine group was significantly higher (t = 3.58, P <0.01). Conclusion Bromocriptine can effectively reverse the multidrug resistance of liver cancer by inhibiting the function of P-glycoprotein.