Study Of NLRC3 In Tumorigenesis Of HCC And Cancer Therapy Of Mineralized Adenovirus

Hepatocellular carcinoma(HCC)is one of the most common causes of cancer-related deaths worldwide.Particularly in China,it accounts for over 50%of all HCC cases in the world due to high rate of chronic hepatitis B virus(HBV)infection.Despite the fact that advanced modern medicine promotes the development of techniques for the diagnosis and therapy of HCC,the prognosis and survival of HCC patients remain disappointing because of high rate of recurrence and metastasis.Thus it is important to elucidate the potential molecular mechanism of the development of HCC for finding effective methods of diagnosis and therapy.Recently,gene therapy technology has developed rapidly as a method for introducing exogenous therapeutic genes into cancer cells.However,the safety and effectiveness is still an urgent problem.In this paper,we will explore the molecular mechanism in the progress of HCC and mineralized oncolytic virus mediated target therapy was also performed.The main contents and results of this paper are as follows:In this study,we first studied the expression of NLRC3 in HCC and analyzed its correlation with clinicopathological features,and the effect of NLRC3 on the biological behavior of HCC cells in vitro was further explored.In order to study the targeted gene therapy for HCC,we introduced the mimetic nucleoprotein W6p into the capsid protein of oncolytic adenovirus as the nucleation site of calcium phosphate mineralization,and oncolytic adenovirus was biomineralized under physiological conditions.Carrying both TRAIL and Smac genes,mineralized virus was constructed target HCC sites using EPR effect of solid tumors,and play a dual gene oncolytic role.Aanalysis based on bioinformatic data showed that high levels of NLRC3 mRNA correlated with a favorable prognosis.Furthermore,expression of NLRC3 was significantly reduced in tumors the compared to noncancerous hepatic tissues,suggesting that NLRC3 potentially modulate tumorigenesis.The Edmondson grades and tumor metastasis reciprocally correlated to NLRC3 levels.Kaplan-Meier survival analysis revealed that HCC patients with high expression of NLRC3 have a more favorable prognosis than those with low expression of NLRC3.We then used shRNA to knock down NLRC3 expression in HCC cell lines and evaluated its effect on cell proliferation and apoptosis.Suppression of NLRC3 expression promoted cell proliferation and inhibited apoptosis in vitro.This suggested to us that NLRC3 may play as a tumor suppressor in HCC and its decreased expression may be used for the identification of tumor metastasis and progression in HCC.Biomineralized oncolytic adenovirus CaP@W6p-ZD55 was found to be a relatively homogeneous nanoparticle with a particle size of 50-80 nm,which has good biocompatibility.CaP@W6p-ZD55 can significantly increase the infection efficiency of adenovirus,especially in cells with low levels of the attachment receptor,Coxsackievirus and adenovirus receptor(CAR),expression.Thus,it can be inferred that CaP@W6p-ZD55 can be absorbed into cells through non-specific pathways assisted by mineral phase.The inorganic shell can protect the adenovirus and does not stimulate the body to produce anti-virus antibodies,nor is it inactivated by antibodies.After treatment by calcium phosphate mineralization,the thermal stability of the virus was significantly enhanced.The further accelerated inactivation experiment showed that the inactivation rate of CaP@ZD55 was slower than that of the non-mineralized virus with the increase of temperature.In vivo experiments showed that CaP@W6p-ZD55-TRAIL-IETD-Smac could increase the number of viruses accurately reaching the tumor site,inhibit the growth of transplanted tumor and prolong the survival time of mice bearing tumor,which enhanced anti-cancer effect.In conclusion,our data showed that the low expression of NLRC3 in HCC is significantly related to poor prognosis.Its downregulated expression also promotes proliferation and inhibits apoptosis of HCC cells in vitro,suggesting that NLRC3 may play a tumor suppressor role in the occurrence and development of HCC.Further study of the mechanisms may provide new strategies for the prevention and treatment of HCC.The oncolytic adenovirus biomineralized by calcium phosphate has the characteristics of good biocompatibility and thermal stability,while avoiding the combination of antibodies and increasing the efficiency of cell infection.Mineralized oncolytic adenovirus carrying TRAIL-IETD-Smac fusion gene can significantly inhibit the growth of tumor,especially in tumor with low expression of CAR.Biomineralization based on calcium phosphate may provide a new way for oncolytic adenovirus-mediated gene therapy.