The Apoptosis Effects Of 2-deoxyglucose（2-dg） Rh₂ On CervicalCancer Siha Cells In Vitro

Cervical cancer is one of the most serious challenges for women’s health.In recent years,the rapid development of Chinese society,as an ageing population,food safety issues,as well as the destruction of the natural environment,and many other factors,the morbidity and mortality of cervical cancer is gradually rising.According to the current studies,the main method of medical treatment for cervical cancer is Surgery,radiotherapy,chemotherapy,concurrentchemoradiotherapy,and immunotherapy.Clinically,the treatment of cervical cancer is mainly surgical treatment.Other treatment methods are complementary,the comprehensive treatment effect is the best.However,tumor cells can produce drug resistance and toxic side effects of chemotherapy drugs.The drugs cause severe adverse reactions in the treatment of cervical cancer patients.Therefore,a novel anti-tumor drug with high efficiency and low toxicity is sought to improve the clinical efficacy of cervical cancer.Reducing mortality and improving the quality of life of patients has become one of the hot spots in recent years.Ginsenoside is one of the important bioactive constituents of ginseng.Because of its antitumor activity.It has been studied for many years.The antitumor activity of ginsenoside Rh₂ monomers is the strongest in the numerous subtypes of ginsenosides.It has the effect of inhibiting tumor cell proliferation and inducing apoptosis of tumor cells.2’-deoxyglucose（2’-dg）is also known as 2-mannose and deoxymannose.It also possesses the pharmacological action of inhibiting tumor cell and proliferation,and can be prepared into novel anti-tumor new drugs to provide new ideas for clinical treatment.This experiment will study the effects of 2’-deoxyglucose Rh₂ on the apoptosis of cervical cancer.The effects of 2’-deoxyglucose Rh₂ on the proliferation and mobility of cervical cancer Siha cells are detected by MTT and Transwell cell migration.Mitochondrial membrane potential detection of 2’-deoxyglucose Rh₂ on the apoptosis of Siha cells in cervical cancer.Flow cytometry detection of 2’-deoxyglucose Rh₂ on the cell cycle and apoptosis of cervical cancer cells.The regulation of the expression of cyclin-d1,bcl-2 and Bax protein in 2’-deoxyglucose Rh₂ was detected by western-blot method.The results of MTT assay showed that at 48h,the 2’-deoxyglucose-Rh₂（50μg/mL）group was significantly different from the 37.5μg/mL group at the same time（P<0.05）,and compared with the same concentration at 24 h and 72 h group.The difference was statistically significant（P<0.05）,indicating that 2h-deoxyglucose（Rh₂）at 48 h and a dose of 50μg/mL had the most significant effect on the proliferation of cervical cancer Siha cells.The results of migration chamber experiments showed that the effect of 2’-deoxydeoxyglucose Rh₂ 50μg/m L on the migration of cervical cancer Siha cells at 48 h after treatment was significantly different from that of the normal control group（P<0.01）,compared with the 37.5μg/mL group.There was a statistically significant difference（P<0.05）.Mitochondrial Membrane Potential Assay for Apoptosis of Cervical Cancer Siha Cells by 2’-Deoxydeoxyglucose-Rh2.The results confirmed that at 48h,the 2’-deoxydeoxyglucose-Rh₂（50μg/mL）group could induce apoptosis of cervical cancer Siha cells significantly,compared with the normal control group（P<0.01）,and 37.5μg/mL group There was a statistically significant difference（P<0.05）.The effect of 2’-deoxy-glucose-Rh₂ on the cell cycle and apoptosis of cervical cancer cells Siha was detected by flow cytometry.The results showed that at48 hours,at the S phase,the 2’-deoxyglucose-Rh₂（50μg/mL）and（75μg/mL）groups were significantly different from the normal group（P<0.01）,and there was statisticallysignificantcomparedwith37.5μg/mLDifferences（P<0.05）,2’-deoxyglucose,Rh₂（50μg/mL）and（75μg/m L）groups were not statistically significant（P>0.05）.It was shown that 2’-deoxyglucose-Rh₂（50μg/mL）can significantly prevent the progression of S phase,thereby inhibiting the DNA synthesis of cervical cancer Siha cells.The apoptosis rate and apoptotic rate were detected by flow cytometry,which were 2.93%in normal group,28.31%in positive control group,3.43%in 2’-deoxyglucose-Rh₂（37.5μg/mL）,and in 2’-deoxyglucose（50μg/mL）22.81%,2’-deoxyglucose Rh₂（75μg/mL）19.92%.Western-blot results showed that the expression of Cyclin-D1 protein in 2’-deoxyglucose-Rh₂ group was significantly lower than that in normal group（P<0.05）;2’-deoxyglucose-Rh₂（50μg/m L）The level of Bcl-2 protein decreased and the expression of Bax increased in the group,which was statistically significant（P<0.05）.It was shown that 2’-deoxyglucose-Rh₂significantly down-regulated the expression of Cyclin-D1 and Bcl-2 protein and up-regulated the expression of Bax protein.In summary,2’-deoxyglucose-Rh₂ can inhibit the proliferation of cervical cancer Siha cells.On the one hand,it may down-regulate the expression of Bcl-2 protein and up-regulate the expression of Bax protein,thereby reducing mitochondrial membrane potential and inducing apoptosis;On the other hand,the downregulation of Cyclin-D1 protein expression may inhibit the progression of cervical cancer Siha cells.