The Inhibition Of RhoC On SKOV3Growth In Animal Model

Ovarian cancer is a common cancer of the female reproductivesystem,Serious harm to the health of the majority of female compatriots.At present, clinical commonly used treatment mainly with surgery way toremove a malignant tumor cells, and combined with radiotherapy,chemotherapy, etc. Although there is a wealth of clinical experience as abasis, a portion of the ovarian cancer patient’s condition has been somerelief, however, the early diagnosis of ovarian cancer, the effects oftreatment, and relapse rate after treatment with5-year survival rate is notsatisfactory. The moment, gene therapy has become a new field ofexploration cancer treatment. RhoC gene is a member of Rho（Ras-homologous）subfamily proteins,the high expression in ovariancancer, and has been recognized by the majority of experts and scholars.Lot of research have confirmed that the invasive ability of RhoC geneexpression in ovarian cancer and cancer of the positive correlation. Wealso found that RhoC gene has an extremely important role in thedevelopment of ovarian cancer cells in vitro in early. RhoC-miRNAsilencing of eukaryotic expression vector we build successful transfectedSKOV3cells,we found that transfected cells significantly slower growthrate, and the situation of apoptosis was significantly increased, with a further indication of the performance of RhoC gene oncogene in vitrocellular level. Order to further investigate the role of being silenced RhoCgene in vivo situation,in this study we have specific lentiviral vectortransfected into SKOV3cells,and injected into mice observe its effectsituation,to investigate the specific inhibition of RhoC gene expression onovarian cancer animal model.Looking forward to a new method of genetherapy for ovarian cancer provides a scientific basis.Materials and Methods:Select female BALB/c nude mouse,6-8weeks old, weighing18~22g were reared. The packed Lenti-shRhoC and Lenti-NC lentiviralvector commissioned the Central Laboratory of Public Health, JilinUniversity to prepare a virus solution and transfected into logarithmicphase of SKOV3cells. The BALB/c nude mouse were randomlydivided into three groups, the transfected cells were not blank controlgroup transfected with Lenti-NC cells transfected with the negativecontrol group and a group-specific interference Lenti-shRhoC cells, each10. Packet in the subcutaneous injection of1ml (5×106/ml) of SKOV3,Lenti-N, Lenti-shRhoC cell solution. Every two days the mice wereobserved and recorded tumor, the mice were weighed on the measuredand tumor volume was calculated. After inoculation, the mice weresacrificed on the20th, observe gross changes in tumor tissue. Preparationof paraffin sections were stained with HE morphological changes in tumor cells. RhoC-miRNA detection after injection of lentiviral proteinexpression in tumor tissues using Westernblot approach.The result:1.The three groups of mice were intraperitoneally transplanted tumormass and volume changes.20d after injection, mice were sacrificed.Specific interference group(ie, Lenti-shRhoC group) intraperitoneally transplanted tumor formationrate, tumor mass, tumor volume was significantly lower than all of thenegative control group (ie, Lenti-NC group) and control group (ieSKOV3group), P <0.01, with statistical significance. But there was nonegative control group and the control group statistically significant, P>0.05.2.Western blotting detection xenograft mouse peritoneal proteinexpression of RhoC Western-blot test results clearly show method,specific interference group, the negative control group and the controlgroup RhoC protein corresponding values were0.3865±0.0328,0.4728±0.0257,0.0964±0.0164, the result shows that the specific interferencegroup was significantly inhibited RhoC expression in tumor tissuesprotein.3. Human ovarian cancer cells nude mice subcutaneous plant tumorpathological observation Intermediate tumor tissue dissection under cut,visible tumor cells in mouse ovarian tumor tissue blank control group and negative control group were transplanted tumors showed a hump-shapedor leafy growth, slightly harder texture, gray cut surface. After HEstaining, visible under the microscope xenografts of human ovariancancer cell shapes and sizes, and a large stained nuclear atypia andmitotic obvious pathology to determine the organization of malignanttissue.Conclusion:In vivo studies using a lentivirus-mediated RNAi technologyspecific inhibition of RhoC gene in a mouse model of ovarian cancershows: RhoC-miRNA lentiviral injection of human ovarian cancerxenografts in mice intraperitoneally with a tumor volume reduction andinvasion dECREASED; RhoC-miRNA can inhibit RhoC gene expressionlevel.