Proliferation Inhibition Of Rosiglitazone On Human Hepatocellular Carcinoma SMMC7721Cell Xenografts In Nude Mouse

Objective: To investigate the effects of Rosiglitazone(ROZ) on proliferation ofhuman hepatocellular carcinoma cell line SMMC7721in nude mice and its molecularmechanism.Methods:1. Establishment and group of human hepatocellular carcinoma SMMC7721cell linexenografts in nude mouse model: BALB/C mouse were transplanted by ipinjection with1×107SMMC7721cell respectively. A total of20nude mousewere randomly divided into4groups: normal saline group,5-Fu control group,ROZ therapeutic group,5-Fu and ROZ therapeutic alliance group. After the tumorformed, normal saline group was injected with NS;5-Fu control group andtherapeutic groups were injected with drag through abdominal cavity respectively.2. The evaluation of ROZ was detected by human hepatocellular carcinomaSMMC7721cell line xenografts in nude mouse model:Transplant tumor’s weightand volume were observed; Morphologic change was analyzed by opticalmicroscope; Phase distribution of cell cycle was analyzed by flow cytometry(FCM); The expression of PPARγ、Cyclin D1、p21WAF1/Cip1、phosphorylatedRb and total Rb protein were observed by western blot in different groups.Results:1. Human hepatocellular carcinoma SMMC7721cell xenografts in nude mousemodel was successfully established. The volume and weight of transplant tumor of5-Fu control group, ROZ therapeutic group,5-Fu and ROZ therapeutic alliancegroup were significantly decreased than that of normal saline group (P＜0.05),and the antineoplastic rate was50.1%,48.7%,66.4%, respectively, which showed ROZ could markedly inhibite the growth of transplant tumor. The weight oftransplant tumor of normal saline group,5-Fu control group, ROZ therapeuticgroup,5-Fu and ROZ therapeutic alliance group was3.071g,1.512g,1.576g,1.033g respectively.2. In the treated group, the cancer nest was distinct and the size of cancer was fairlyunity, cellular atypia and the pathology karyokinesis were reduced, nuclear andpartial cell organs manifested retrograde alters and partial cells manifest apoptosismorphologic alters, there were a lot of hemorrhage and cellular necrosis in thecancer nest, which are observed by optical microscope.3. Flow cytometry analysis revealed that the cell cycle of SMMC7721cells werearrested in G1phase and higher hyodiplod peak was found(P＜0.05) after treatedwith ROZ or5-Fu in vivo. In the treated groups, the proportion of cells in G1phase were increased markedly compared with normal saline group(P＜0.05),while the proportion of cells in G2phase were decreased and the proportion ofcells in S phase effected a little.4. Western blot analysis confirmed, there were the expression of PPARγ in everygroup, but after treated by ROZ or5-Fu, the expression of PPARγ were increasedmarkedly（P＜0.05）. Western blot analysis also indicated that expression ofCyclin D1declined, p21WAF1/Cip1reinforced, ROZ or5-Fu effectivelydownregulated phosphor-Rb prorein levels（P＜0.05）, while total Rb proteinlevels were not affected.The change of the all protein were the most significant inthe5-Fu and ROZ therapeutic alliance group, and there was no significantdifference among ROZ therapeutic group and5-Fu control group (P＞0.05).Conlusion1. ROZ can significantly inhibit proliferation of human hepatocellular carcinomaSMMC7721cell in nude mice.2. ROZ can induce SMMC7721cell blocked in G1and its apoptosis.3. The effect of growth inhibition and the cell cycle arresting of ROZ in humanhepatocellular carcinoma SMMC7721cell line probably relate to inducedexpression of PPARγ, downregulated expression of Cyclin D1, upregulated expression of p21WAF1/Cip1, and downregulated phosphor-Rb prorein levels.