阿伐麦布中断硫代乙酰胺诱发的大鼠胆管上皮癌变

Avasimibe interrupts cholangiocarcinoma induced by thioacetamide in rats

目的探索阿伐麦布在预防化学致癌物硫代乙酰胺所致胆管上皮癌变中的作用。方法用含300 mg/L硫代乙酰胺的饮用水饲养雄性SD大鼠,3个月后停止饮用并分为两组(n=5):对照组和阿伐麦布干预组。对照组更换为普通饮用水,阿伐麦布干预组用含阿伐麦布(30 mg/kg)的饮用水灌胃饲养。干预3个月后处死所有大鼠,用H-E染色观察大鼠肝脏病变情况,用免疫组织化学染色检测大鼠肝脏病变组织中醛固酮类还原酶家族1成员C1(AKR1C1)和Ki-67的表达。用不同浓度(0、10、20μmol/L)的阿伐麦布处理胆管癌细胞QBC939,用CCK-8法检测细胞增殖能力,用qRT-PCR和蛋白质印迹分析检测QBC939细胞中AKR1C1的表达情况。结果H-E染色结果显示,对照组大鼠均形成了镜下可见的肿瘤,成瘤率为100%(5/5);阿伐麦布干预组仅1只大鼠形成了肿瘤,成瘤率为20%(1/5)。免疫组织化学染色结果显示,阿伐麦布干预组大鼠的肝脏病变组织中AKR1C1和Ki-67的表达水平均较对照组下降(P<0.05,P<0.01)。CCK-8法细胞增殖能力检测结果显示阿伐麦布能够抑制QBC939细胞的增殖。qRT-PCR和蛋白质印迹分析结果显示,经阿伐麦布干预的QBC939细胞中AKR1C1 mRNA和蛋白的表达水平均较对照组下降(P均<0.01)。结论阿伐麦布能够中断化学致癌物所致的肝内胆管癌的发生,是极具潜力的预防胆管癌的药物;AKR1C1是胆管癌潜在的治疗靶点。

Objective To explore the effect of avasimibe in preventing cholangiocarcinoma induced by thioacetamide,a chemical carcinogen.Methods Male SD rats were given drinking water containing 300 mg/L thioacetamide and stopped drinking after 3 months,and then were divided into two groups(n=5):control group and avasimibe intervention group.The control group was given normal drinking water,while the avasimibe intervention group was given drinking water containing avasimibe(30 mg/kg)by gavage.After 3 months of intervention,all rats were sacrificed and liver lesions were observed by hematoxylin-eosin(H-E)staining.The expression of aldo-keto reductase family 1 member C1(AKR1C1)and Ki-67 in liver lesions of rats was detected by immunohistochemical staining.Meanwhile,different concentrations(0,10,20μmol/L)of avasimibe were used to treat the cholangiocarcinoma cell line QBC939,the cell proliferation ability was detected by cell counting kit 8(CCK-8)assay,and the expression of AKR1C1 in QBC939 cells was detected by qRT-PCR and Western blotting.Results The results of H-E staining showed that all rats in the control group developed visible tumors under the microscope,and the tumor formation rate was 100%(5/5),while only one rat in the avasimibe intervention group developed tumors,with a 20%(1/5)tumor formation rate.The results of immunohistochemistry showed that the expression levels of AKR1C1 and Ki-67 in liver lesions of rats in the avasimibe intervention group were significantly lower than those in the control group(P<0.05,P<0.01).The results of CCK-8 assay showed that avasimibe significantly inhibited the proliferation of QBC939 cells.The results of qRT-PCR and Western blotting showed that the expression levels of AKR1C1 mRNA and protein in QBC939 cells after the intervention of avasimibe were significantly decreased compared with those of the control group(both P<0.01).Conclusion Avasimibe can interrupt the occurrence of intrahepatic cholangiocarcinoma caused by chemical carcinogens and is a potential drug for preventing cholangiocarcinoma.AKR1C1 might be a potential therapeutic target of cholangiocarcinoma.