Study On The Mechanism Of Dihydroartemisinin In The Prevention And Treatment Of Cervical Lesions

Objective:Cervical cancer is the fourth most common female malignancy in the world and a major health challenge worldwide.The 2015 survey found that about 270,000cervical cancer patients died in low-and middle-income countries,and the mortality rate in these countries is 18 times that of developed countries.In recent years,the incidence of cervical cancer in China has also become younger and rising.Although the coverage of cervical cancer screening has been expanding in recent years,treatments have progressed with technological innovation.However,there are still large individual differences and limitations in the accuracy of diagnosis of HPV infection,cervical precancerous lesions and cervical cancer and the effectiveness of treatment.Artemisinin（ARS）has long been known as an effective antimalarial drug,and its research shows that it is anti-cancer,anti-viral,anti-parasitic,anti-fungal,allergic inflammation,specific inflammation,infectious inflammation and leukemia.The treatment has a significant effect.Therefore,this study is based on the anti-tumor properties of dihydroartemisinin,to determine its role in cervical cancer cells and its mechanism.Methods:1.Collect cervical inflammatory tissue,low-grade squamous intraepithelial lesion（LSIL）,high-grade squamous intraepithelial lesion（HSIL）,cervical squamous cell carcinoma（Cervical squamous cell carcinoma）a total of 172 cases.Immunohistochemistry was used to detect the expression levels of ki67 and p16 in different grades of cervical lesions.2.Using cervical cancer HeLa cell line as an ex vivo model of cervical cancer cells,CCK-8 uses cell activity and cytotoxicity assay kit（Cell Counting Kit-8,CCK-8）to determine dihydroartemisinin in vitro.The effect of50%inhibition of cervical cancer cells（Half maximal inhibitory concentration,IC50value）,and the drug’s ability to increase the value of cervical cancer cells.The expression levels of apoptosis-related proteins and migration-related proteins in cells were detected by western blot.The effects of dihydroartemisinin on the migration of cervical cancer cells were examined by cell scratch assay.Western blot was used to detect the expression of ki67 and p16 in HeLa cells after drug treatment,and it was confirmed whether artemisinin has anti-HPV infection.Results:1.The positive expression rates of ki67 and p16 in different cervical tissues were as follows:the positive expression rate of p16 in the inflammation group was11.11%（4/36）,and the positive expression rate in the low-grade cervical intraepithelial neoplasia group was 60.00%（30/50）,the positive expression rate of high-grade cervical intraepithelial neoplasia group was 94.34%（50/53）,and the positive expression rate of cervical cancer group was 100.00%（33/33）.The positive expression rate of ki67 in the inflammation group was 30.56%（11/36）,and the positive expression rate in the low-grade cervical intraepithelial neoplasia group was 56.00%（28/50）.The positive expression rate of the high-grade cervical intraepithelial neoplasia group.At94.34%（50/53）,the positive expression rate of cervical cancer group was 100.00%（33/33）.The positive expression rates of p16 and ki67 in different grades of cervical lesions were compared（χ₁²=87.474,P₁<0.001;χ₂²=61.477,P₂<0.001）.The expression rates of p16 and ki67 in different cervical tissues were different.The positive expression rate of P16 and ki67 was positively correlated with the grade of cervical tissue lesions.2.The drug effects of dihydroartemisinin on HeLa cells are mainly as follows:1 The experimental results of CCK-8 cell proliferation activity suggest that artemisinin can inhibit the proliferation of cervical cancer cells in a time-and concentration-dependent manner.The IC50 value of artemisinin for HeLa cells was calculated to be22.86±3.765umol/l.2DHA promotes apoptosis of HeLa cells through mitochondria and endoplasmic reticulum pathways.The expression of pro-apoptotic proteins clever-Caspase3,clever-Caspase9 and clever-Caspase12 in the mitochondrial pathway increased,and the expression of Bcl-2 and Bcl-xl was decreased.The expression of related protein Chop increased in the endoplasmic reticulum stress pathway.3 cell scratch experiments showed that DHA inhibited the migration of cervical cancer cells.After treatment with dihydroartemisinin,the expression of E-cadherin in HeLa cells increased.4 Artemisinin resists malignant proliferation of cells caused by HPV infection.After 36 hours of drug treatment,the expression of ki67 protein in HeLa was significantly decreased and the expression of p16 protein was increased.Conclusions:Dihydroartemisinin has a significant inhibitory effect on the occurrence and development of cervical cancer,and has broad clinical application prospects,and can be used as a new weapon for future prevention and treatment of cervical cancer.