Effect Of Pirarubicin Collaborate With HepaCAM Gene On Proliferation In Bladder Cancer Cell

Part one: The chemotherapeutic efficacy of hepaCAM on bladdercancer cells BIU-87treated with pirarubicin.Objective: To explore the chemotherapeutic efficacy of bladdercancer cell BIU-87treated with the adenovirus mediated liver cell adhesionmolecules (hepatocyte cell adhesion molecule, hepaCAM) and Pirarubicin.Methods: RT-qPCR was used to test the mRNA levels of cyclinD1and CDK2in the pAdH5-hepaCAM treatment groups, the pirarubicintreatment groups, pAdH5-hepaCAM collaborate with pirarubicin treatmentgroup and uninfected group of bladder cancer BIU-87. Cell MTT assay wasused to observe the inhibition rates of cells in different groups. FCM wasused to analyze the changes of cell cycle.Results: The result of real-time PCR showed that the mRNA levels ofcyclinD1and CDK2in pAdH5-hepaCAM collaborate with pirarubicintreatment group was significantly lower than other groups.(p <0.05);MTTassay showed inhibition rate in pirarubicin/pAdH5-hepaCAM group wasmarkedly increased as compared with that in pirarubicin group, pAdH5-hepaCAM group and control group;In FCM assay,the cell inG0/G1was significantly increased in pirarubicin/pAdH5-hepaCAMgroup than control group， pirarubicin group and pAdH5-hepaCAMgroup(p<0.05).Conclusion: Cell proliferation was inhibited, cell cycle arrest at theG0/G1phase in BIU-87cell treated with pirarubicin and pAdH5-hepaCAMcompared with control group and Separate treatment groups.Our resultsshow that pirarubicin collaborate with hepaCAM gene can enhance theeffect of chemotherapy on bladder cancer. Part tow: The effect of hepaCAM gene on the growth of bladdercancer in vivoObjective: The effect of hepaCAM on the growth of bladder cancer innude mouse injected with BIU-87cell was detectedMethods:10nude mice were randomly divided into the control andhepaCAM group.1*107BIU-87cells were inoculated in right lowerextremity dorsolateral subcutaneous in a nude mouse. After injection thenude mice were still cultivate with SPF environment, observe tumor casesand record tumors volume and the weight of nude mice every day. After three weeks, the mice were killed. Compared with the tumor size in twogroups of nude mice then immunohistochemical was used to detect theexpression of hepaCAM.Result：The tumor size of control group was significantly larger thaninfection group. The result of immunohistochemistry showed that hepaCAMprotein expression in control group was significantly lower than infectiongroup.Conclusion: Vivo experiments show hepaCAM gene cansignificantly inhibit tumor growth using adenovirus vector and furthervalidate hepaCAM gene is tumor suppressor gene of bladder cancer.Therefore，the hepaCAM gene with adenovirus vector is a new effectivemethod for gene therapy, which can provide an important basis for futureclinical research and treatment.