| Objective1.By observing the clinical effect of "external Chinese medicine prescription" in the treatment of persistent cervix HR-HPV infection,the TCM dialectical thinking and rational prescription for the effective treatment of cervix HR-HPV infection were explored.2.To investigate the effect of "external prescription" on the proliferation and apoptosis of cervical cancer Siha cells,and to analyze the mechanism of tumor inhibition.3.To study the antitumor effect of "external prescription of traditional Chinese medicine" on cervical cancer in nude mice,and to study its toxic reaction to nude mice.4.To explore the effect of "external Chinese medicine" on the expression of cancer protein associated with cervical cancer,and to analyze the mechanism of tumor inhibition.Methods1.A total of 70 patients with HR-HPV infection of cervix cervix with dampness and heat were randomly divided into treatment group(35 cases)and blank control group(35 cases).The course of treatment was set at 3 months and the curative effect was evaluated after 3 months of continuous treatment.The clinical symptom score and HPV negative rate before and after treatment were compared between the two groups to evaluate the curative effect.2.Human cervical cancer Siha cells in logarithmic growth phase were treated with different concentrations of 15 mg/ml～7.5 mg/ml～3.25 mg/ml,and Siha cells were used as control group to detect cell growth inhibition.Trypan blue staining was used to detect cell viability and HOECHST 33342/P1 staining was used to detect the morphological changes of apoptotic cells.3.A nude mouse model of human cervical cancer Siha cells bearing tumor was established.The nude mice were randomly divided into three groups:high dose group,middle dose group,low dose group,cisplatin group,blank control group,tumor size and weight of nude mice,which were divided into three groups:high dose group,middle dose group,low dose group,cisplatin group,blank control group,daily measurement of tumor size and weight of nude mice.The intake of water was observed for 28 days.Nude mice were killed 24 hours after drug withdrawal and tumor tissues were taken out for experimental study.4.The tumor tissues were stained with HE and immunohistochemical SP method was used to detect the expression of E6E7 RB and p53 protein in the tumor tissue.Results1.In the blank control group,there was no statistical difference in the scores of clinical symptoms of TCM before and after treatment,and the scores of clinical symptoms of TCM in the treatment group were significantly lower than those before and after treatment,and the scores of clinical symptoms of TCM in the treatment group were significantly lower than those before and after treatment.There was significant statistical difference between the two groups in terms of P<0.01,and the difference between the two groups before and after treatment indicated that the treatment group of external Chinese medicine was superior to the blank control group in reducing the score of clinical symptoms of TCM(P<0.01).After 4 months and 12 months of treatment of HPV infection,the total effective rate of the treatment group was 54.29%and 65.72%respectively,which was better than that of the blank control group.The total effective rate of the two groups was significantly higher than that of the control group(P<0.01).2.MTS assay showed that the growth of cervical cancer SiHa cells was inhibited in different degree in each dose group of "external Chinese medicine prescription".There were significant differences between the 3.125 mg/L dose group and the blank group(P<0.05),IC50 was 15±2.1 mg/mL,and trypan blue staining:"external Chinese medicine prescription" was used to detect the growth of cervical cancer SiHa cells in different degrees,and there was a significant difference between the 3.125 mg/L group and the blank group(P<0.05).The growth of Siha cells was inhibited to some extent.The inhibition rate of cell growth in each concentration group was higher than that in the control group(P<0.05),the intercellular quality of Siha cells was loosened,cytoplasmic granules increased,chromatin condensed,partitioned,cytoplasm concentrated,and HOECHST 33342/PI staining.It was observed under fluorescence microscope that "external prescription of traditional Chinese medicine" could induce apoptosis(irregular morphology of cells,bright blue fluorescence from early apoptotic cells,fragmentation of some apoptotic cells in the shape of dots and different sizes).3.The volume of transplanted tumor in the "external prescription" group was significantly smaller than that in the blank control group(P<0.05).The tumor inhibition rates of the high,middle and low dose groups were 36.233.24%and 14.94%respectively.The results showed that there were obvious necrotic changes in the transplanted tumor tissue of the nude mice in the "external Chinese medicine prescription" group.4.Compared with the blank control group,the expression level of E6 and E7 protein in the high and middle dose groups were significantly lower than that in the control group.Compared with the blank control group,the expression of RbP53 protein in the high dose and middle dose groups of Chinese herbal prescription was lower than that in the blank control group,and the difference was statistically significant.Conclusion1.The effect of external use of "Chinese traditional Chinese medicine" in the treatment of the persistent infection of HR-HPV in the patients with damp-heat and lower injection can obviously improve the rate of HPV transfer,and can obviously improve the clinical symptoms of the Chinese medicine.Therefore,the external use of the traditional Chinese medicine prescription is worthy of being applied to the infection of the cervical HR-HPV of the damp-heat-injection type cervix.2."External traditional Chinese medicine" can inhibit the proliferation of cervical cancer Siha cells and induce apoptosis.3."External traditional Chinese medicine" has obvious inhibiting effect on the growth of human cervical cancer Siha cell transplantation tumor.4.The expression of Rb and P53 proteins can be regulated by "Chinese traditional Chinese medicine",and the expression of E6 and E7 proteins is downregulated.The expression of Rb and P53 proteins is regulated,and the expression of E6 and E7 proteins is downregulated,which is one of the mechanisms for inducing apoptosis of cervical cancer cells. |
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