Matrine Inhibited The Proliferation Of Cervical Cancer Cells By Regulating The Expression Of MiR-375-3p And HPVE6 And HPVE7

Objective:In this study,the inhibitory effect of matrine on Siha and Caski cells of cervical cancer was investigated to explore the effect of matrine on the expression of mir-375-3p and HPVE6 HPVE7 in Siha and Caski cells of cervical cancer.In addition,we try to further clarify the effect of matrine on the prevention and treatment of cervical cancer by down-regulating the expression of HPV virus,so as to provide theoretical and experimental basis for the prevention and treatment of cervical cancer with traditional Chinese medicine.Methods:1.The effect of matrine on Siha and Caski cell proliferation in cervical cancer was determined by MTT colorimetry at different time points.2.Flow cytometry was used to detect the cycle of matrine on Siha and Caski cells of cervical cancer.3.The effect of matrine on proliferation inhibition of Siha and Caski cells in cervical cancer was examined by 3.EDU assay.4.The expression of matrine on HPVE6,HPVE7,P16,C-MYC and tumor suppressor genes P21 and P27 in Siha and Caski cells of cervical cancer was detected by WB.5.Rt-PCR was used to detect the effect of matrine on mir-375-3p,so as to verify the effect of mir-375-3p on the expression of HPVE6 and HPVE7,and further inhibit the expression of P21 and P27.6.HPVE6 overexpression plasmid transfection was performed on cervical cancer cells Siha and Caski to test whether HPVE6 overexpression had an effect on P21 and whether it had a reversal effect after matrine drug intervention.7.Animal experimental study on the antitumor effect of matrine monomer.The nude mice were divided into blank group and drug administration group.The nude mice in the drug administration group were injected with matrine solution intraperitoneally every day.After 15 days,the tumor volume and weight of nude mice were observed.The samples were collected and detected by WB and PCR.Results:1.MTT results showed that compared with the blank group,matrine significantly inhibited the proliferation of Siha and Caski cells in cervical cancer(P<0.05).2.Flow cytometry cycle results showed that compared with the blank group,matrine group blocked the cycle of Siha and Caski cells in the G0/G1 phase of cervical cancer,and the difference was statistically significant(P<0.05).3.The results of 3.EDU experiment showed that compared with the blank group,matrine effectively reduced the proliferation of Siha and Caski cells in cervical cancer,with statistically significant difference(P<0.05).4.WB results showed that compared with the blank group,matrine group up-regulated the protein expression of P21 and P27 in Siha and Caski cells of cervical cancer,and down-regulated the protein expression of HPVE6,HPVE7 and P16,with statistically significant differences(P<0.05).5.PCR results showed that compared with the blank group,matrine up-regulated the expression of mir-375-3p in Siha and Caski cells of cervical cancer(P<0.05).6.Siha and Caski cells were transfected with hpv-e6 overexpressed plasmids.The results showed that HPVE6 protein was up-regulated and P21 protein down-regulated in the HPVE6 transfection group compared with the blank group and the blank plasmids group.P21 was significantly up-regulated in HPVE6 transfection group after administration.The results of animal experiments on the antitumor effect of matrine monomer showed that compared with the blank group,the volume,weight and in vivo imaging of the tumor of nude mice in matrine administration group were significantly reduced.The tumor tissues were detected by WB and PCR.The results of WB showed that HPVE6 and HPVE7 were significantly down-regulated and P21 was significantly up-regulated.The PCR results showed that mir-375-3p was significantly up-regulated in the drug administration group compared with the blank group.Conclusion:Matrine can inhibit the proliferation of Siha and Caski in cervical cancer.Western blot showed the regulatory effect of matrine on hpvl6-related factors(down-regulated hpv16-e6 and hpv16-e7)and cancer-related factors(down-regulated P21 and P27).P16 and cmyc were up-regulated.It is suggested that matrine may inhibit the occurrence of cell carcinoma by regulating the transcription and expression of human papillomavirus(HPV).In addition,matrine can also up-regulate the expression of mirna-375-3p,thus significantly inhibiting the expression of viral proteins.The overexpression of hpv-e6 reversed the activity of P21 induced by matrine,and the down-regulation of P21 significantly reversed the effect of matrine on cell viability.According to these results,matrine may regulate HPV function by mediating the expression of mirna-375-3p.More importantly,to further demonstrate the antitumor effect of matrine on CC,we compared xenograft tumor models.The results showed that matrine can obviously inhibit the formation of tumor in vivo.The protein levels of HPV E6/E7 in tissues decreased significantly,while matrine significantly increased the expression levels of P21 protein and mirna-375-3p in tissues.These data further elucidated the antitumor activity of matrine.In summary,matrine may regulate the proliferation of CC cells through the expression of mir-375-3p and HPV proteins,thus inhibiting the proliferation of CC cells.These findings may provide new insights into preventing the formation of CC.