Study On The Effect Of Oncolytic Adenovirus Coagulation Factor X Complex Enhancing The Infection Ability Of Virus On Ovarian Cancer Cells And Its Safety

Objective: Ovarian cancer is the gynecological malignant tumor with the highest mortality rate.Platinum resistance and lack of effective drug options are important causes of death.As a strong and effective anti-tumor agent,oncolytic adenovirus plays an important role.However,a low infection rate of oncolytic adenovirus on ovarian cancer cells,and the currently applied local injection method limit its therapeutic effect on ovarian cancer which easily metastasizes to the abdominal cavity.In this study,by combining oncolytic adenovirus with coagulation factor X(FX)to form a complex,the ability of oncolytic adenovirus to infect ovarian tumors and the effect of oncolytic adenovirus on liver damage were explored by intraperitoneal injection,providing a new oncolytic adenovirus protocol suitable for the ovarian cancer treatment.Methods: Two oncolytic adenoviruses,Ad5/d E1A/EGFP and Ad5/d E1A/Luciferase,were constructed by molecular biology methods.The tumor-selective replication of the virus was verified by q PCR.Coxsackie adenovirus receptor(CAR)expression and oncolytic adenovirus infection ability on ovarian cancer cell lines were explored by immunoblotting experiments and virus binding experiments.The infection effect efficiency of oncolytic adenovirus on ovarian cancer cell was explored through premixed FX.That the oncolytic adenovirus premixed with FX enters the cell through the heparan sulfate proteoglycan(HSPG)molecule on the cell surface was verified after treatment with heparinase I.The binding ratio of oncolytic adenovirus and FX was analyzed through binding experiments and flow cytometry.In vivo imaging technique was used to explore the tumor tissue infection and biodistribution of oncolytic adenovirus coagulation factor X complex in the Balb/c mouse model.The influence of the virus on the liver and kidney function of mice is detected by the levels of plasma alanine aminotransferase,aspartate aminotransferase,creatinine and urea nitrogen.Results: The two constructed oncolytic adenoviruses,Ad5/d E1A/EGFP and Ad5/d E1A/Luciferase,all showed the conditional replication of the virus.Most ovarian cancer cell lines have low expression of CAR,and oncolytic adenoviruses have a low infection rate for most ovarian cancer cells.The expression level of CAR on cancer cells is positively correlated with the infection rate(P = 0.0048,r = 0.7013).FX premixing complex enhances the infection efficiency of oncolytic adenovirus on 7kinds of ovarian cancer cells,which has a high transfection efficiency at 1% of physiological concentration.In vitro experiments have proved that adenovirus coagulation factor X complex enters tumor cells mainly through HSPG on the cell surface.In the nude mouse subcutaneous tumor model bearing human ovarian cancer,intratumoral injection of the oncolytic adenovirus coagulation factor X complex can also increase the infection ability of adenovirus on ovarian tumor tissue.In the Balb/c mouse model,after intraperitoneal injection,compared to the single virus group,the oncolytic adenovirus coagulation factor X complex group mainly accumulates in the lower part of the abdominal cavity with a lower liver distribution,and can increase the residence time of adenovirus in the abdominal cavity.The oncolytic adenoviruscoagulation factor X complex can also reduce the effect of oncolytic adenovirus alone group on liver and kidney function and mouse body weight.Conclusion: Combination of the oncolytic adenovirus with coagulation factor X enhance the oncolytic adenovirus’ s ability to infect ovarian cancer cells,and intraperitoneal injection of the oncolytic adenovirus coagulation factor X complex can reduce the liver distribution of oncolytic adenovirus,increasing the residence time in the abdominal cavity and reduce the toxicity,which is expected to be applied to the oncolytic adenovirus treatment strategy for ovarian cancer abdominal metastasis.