miRNA-17-92高表达白血病动物模型化疗敏感性及复方浙贝颗粒增敏作用研究

Chemosensitivity of miRNA-17-92 high expression in leukemia animal models and the chemosensitization function of compound Zhebei granules

目的观察miRNA-17-92高表达对白血病小鼠化疗敏感性的影响及复方浙贝颗粒的化疗增敏洼。方法应用L1210细胞及miRNA-17-92高表达细胞，DBA／2N小鼠按组别尾静脉分别注射细胞100×10-4／只。将造模小鼠分为5组：miRNA-17-92高表达L1210细胞白血病小鼠（miRNA-17-92）观察组、miRNA-17-92化疗组、miRNA-17-92化疗联合复方浙贝组、L1210细胞白血病小鼠（L1210）化疗组、正常对照组。化疗组经腹腔给药，注射环磷酰胺；化疗联合复方浙贝组加用复方浙贝颗粒灌胃。在实验过程中每7d检查1次外周血常规及白细胞分类；于给药第15天或濒死前处死动物，检测外周血常规及白细胞分类、骨髓形态学检查白血病细胞的比例、脾脏称重并观察病理组织学的改变、酶联免疫吸附试验检测P糖蛋白（P-gp）的表达。结果miRNA-17-92观察组、miRNA-17-92化疗组、miRNA-17-92化疗联合复方浙贝组、L1210化疗组、正常对照组给药第15天血红蛋白和血小板计数比较，差异有统计学意义（F=5．76，P=0．002；F=16．90，P=0．002）；miRNA-17．92观察组较正常对照组外周血白细胞计数下降（P〈0．05）；外周血白血病细胞比例分别为（37．2±13．1）％、（5．7±1．9）％、（1．3±0．8）％、（0．8±0．3）％、0，骨髓白血病细胞比例分别为（26．9±11．0）％、（6．8±2．0）％、（3．2±1．0）％、（1．7±1．2）％、0，差异均有统计学意义（F=24．38，P=0．0000；F=19．71，P=0．0000）。脾脏重量分别为（0．70±0．18）、（0．20±0．07）、（0．44±O．17）、（0．23±0．19）、（0．60±0．18）g，差异有统计学意义（F=8．78，P=0．032），化疗可减轻白血病脾脏浸润。P-gP分别为（33±11）、（27±11）、（32±11）、（20±11）、（17±3）ng／ml，差异有统计学意义（F=10．42，P=0．034），复方浙贝颗粒未明显减低P-gP表达。结论miRNA-17-92高表达L1210小鼠化疗反应率减低，复方浙贝颗粒可以提高miRNA-17-92高表达的化疗反应率。

Objective To observe the effect of miR-17-92 overexpression on the chemosensitivity of leukemia mice and the chemosensitivity of compound Zhebei granules. Methods The miR-17-92 over- expressed ceils and L1210 ceils were conventionally infused into each DBA/2N mouse through the vena caudalis at the cellular concentration of 100×10-4. The mice were divided into 5 groups with a randomized method, including miR-17-92 high expression L1210 cell leukemia mice （miR-17-92） observation group, miRNA-17-92 chemotherapy group, miRNA-17-92 chemotherapy combined with compound Zhebei granules group, L1210 chemotherapy group and normal control group. The chemotherapy groups were given cyclophosphamide by intraperitoneal injection, or chemotherapy combined with compound Zhebei granules group plus compound Zhebei. In the course of the experiment, the routine blood and white blood cell classification were checked every 7 days. The animals were killed on the 15th day or before dying. Peripheral blood routine and white blood cell classification were detected, the proportion of leukemia cells was checked by using bone marrow morphology. Furthermore, it was necessary to weigh the spleen and observe histopathological changes. P-glycoprotein （P-gp） expression was detected by using enzyme-linked immunosorbent assay. Results The difference between hemoglobin and platelet count on the 15th day after administration was statistically significant in the miRNA-17-92 observation group, miRNA-17-92 chemotherapy group, miRNA-17-92 chemotherapy combined compound Zhebei group, L1210 chemotherapy group, and normal control group （F = 5.76, P = 0.002; F = 16.90, P = 0.002）; Compared with the normal control group, peripheral blood leukocyte count in the miRNA-17-92 observation group was decreased （P 〈 0.05）. The proportion of peripheral blood leukemic cells was （37.2±13.1） %, （5.7±1.9） %, （1.3±0.8） %, （0.8±0.3） %, 0, and the proportion of myeloid leukemia cells was （26.9±11.0） %, （6.8±2.0） %, （3.2±1.0） %, （1.7±1.2） %, 0, and the difference was statistically significant （F = 24.38, P = 0.000 0; F = 19.71, P = 0.000 0）. The spleen weights were （0.70±0.18）, （0.20±0.07）, （0.44±0.17）, （0.23±0.19）, （0.60±0.18） g respectively, and the difference was statistically significant （F = 8.78, P = 0.032）. Chemotherapy could reduce leukemia spleen infiltration. P-gp was （35±11）, （27±11）, （32±11）, （20±11）, （17±3） ng/ml, and the difference was statistically significant （F = 10.42, P = 0.034）. Compound Zhebei granules did not significantly reduce P-gp expression. Conclusions The overexpression of miRNA-17-92 leads to the decrease of chemotherapy response rate. Compound Zhebei granules combined with chemotherapy could improve the response rate of miR-17-92 high expression L1210 mice.