| Backgroud: Endometrial cancer is one of the three malignant tumors in gynecologic reproductive system,whose incidence rate is second in China.The latent symptoms of endoemtrial cancer in early stage leads to its treatment delay.Nowaday,the current treatment methods for endometrial cancer mainly include surgery,chemotherapy,postoperative or post-radiotherapy adjuvant chemotherapy,radiation therapy,endocrine therapy,and targeted drug therapy which can specifically select carcinogenic sites for combination.Although the treatment methods are constantly updated and improved,the treatment effect is still disappointing.Gene therapy is a new type of treatment method that has been rapidly developed in recent years and has good effects on tumorous diseases and has broad application prospects in future.IL-24 is a tumor suppressor gene with cytokine characteristics and broad-spectrum anti-tumor characteristics.Its proteins can specifically kill tumor cells without having any toxic effect on normal cells,so it may be a promising therapeutic gene for gene therapy.Although many studies including melanoma and breast cancer have shown that IL-24 has anti-tumor effect,so far there is no research on the treatment of endometrial cancer by IL-24.Objective: IL-24 is considered to be a cytokine tumor suppressor gene with broad-spectrum tumor-specific killing effects.However,up to now there is no relevant report on the effect of IL-24 on endometrial cancer.This study is to discover the effect of IL-24 on endometrial cancer and its related molecular mechanism.Methods: Firstly,Oncomine database was searched for the expression differences of IL-24 gene between endometrial cancer patients and normal people,and the relationship between IL-24 gene and clinical characteristics of endometrial cancer patients was explored.Secondly,Immunohistochemical experiments were performed to verify the expression difference of IL-24 protein in endometrial cancer population and normal population.Thirdly,we use Lipo2000 liposome transfection technology to transfect IL-24 plasmid into endometrial cancer cell line which was named Ishikawa.Real-time Quantative PCR Detecting System and Western Blot was used to verify its transfection efficiency.The cell migration test,Transwell test,CCK-8 test,and TUNEL test were used to study the changes in the migration ability,invasion capability,proliferation facility and apoptosis of endometrial cancer cells after overexpression of IL-24,the possible molecular mechanism was determined subsequently.Finally,a nude mouse xenograft tumor experiment was used to study the anti-tumor effect of IL-24 in vivo,and HE staining and immunohistochemical staining of tissue sections were used.Results: We found significantly different expression of IL-24 protein between endometrial cancer patients and control group,among which IL-24 showed a higher expression level in cancer patients than that of the control group.We further transfected IL-24 plasmid in endometrial cancer cell line,then q PCR and WB experiments proved that the overexpression of the cells was successful.The later experiments indecated that the migration ability and invasion capability of the cells were decreased while cell apoptosis was increased when overexpression of IL-24 in Ishikawa cells.Meanwhile,WB results showed that the expression of Bcl-2,MMP-3,VEGF,caspase-9 and Caspase-3 were decreased when IL-24 was overexpressed in Ishikawa cells,while Bax,cytochrome C and the ultimate executor of apoptosis,Cleaved-Caspase-3 were increased.The results of xenograft tumor experiment in nude mice showed that the ability of tumor formation was weakened when IL-24 was over-expressed and the ability of inducing angiogenesis was decreased as well.Conclusion: Our study demonstrates IL-24 can inhibit the development of endometrial cancer.IL-24 can promote the apoptosis of endometrial cancer cells through mitochondrial related endogenous signaling pathway,which indicates that IL-24 may be a promising candidate gene for gene therapy of endometrial cancer. |
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