Detection And Experimental Treatment Of Human Papillomavirus Infection In Cervix

Objectives1.To investigate the applicable value of Hybrid CaptureⅡ(HC-Ⅱ) and the flow fluorescent hybridization assay on human papillomavirus(HPV) detecting.To evaluate the clinical accuracy of the two method for the diagnosis of cervical neoplasia.To detect the most common HPV subtype in different grades of cervical lesions.2.To investigate the mechanism of the proliferative inhibition of HPV16 by curcuma aramatica oil.Methods1.872 patients with abnormal cervical cytology results who accepted cervical cancer screening in China-Japan Friendship Hospital from June 2006 to April 2007 took HR-HPV testing(HC-Ⅱmethod) and all patients underwent colposcopy and biopsy..All patients were divided into five groups as NILM,CINⅠ,CINⅡCINⅢand invasive cervical cancer(ICC).Association between HR-HPV load and CINs were evaluated by Spearman rank correlation.Those with histological pathology diagnosis as cervical intraepithelial neoplasia(CIN)Ⅱor above were HPV subtyped by flow fluorescent hybridization assay.The accordance rate of the two assays was compared. The applicable value on diagnosis of cervical lesions by the two methods was evaluated according to the diagnosis of histological pathology.The infection rates of different genotypes in each group were calculated.2.The human cervical cancer cell line-CaSki and the immortalized cervical epithelial cell-H8 were cultured with different concentration of baofukang suppository The human cervical cancer cell line-CaSki,SiHa and the immortalized cervical epithelial cell-H8 were cultured with different concentration of curcuma aramatica oil.The proliferative inhibition of different concentration of curcuma aramatica oil or baofukang suppository on cell lines was measured by methyl thiazolyl tetrazolium(MTT) assay and flow cytometry assay;The mRNA expression of HPV16 E6E7 was determined by RT-PCR semi-quantitatively.Results 1.The sensitivity,specificity,positive predictable value and negative predictable value for CIN2 by HC-Ⅱwere 93.1%,66.8%,59.9%,94.8%,respectively.There was significant differentce between NILM and CINs or ICC,while no significant difference was observed among CINⅠ,CINⅡ,CINⅢand ICC.The total accordance rate of the flow fluorescent hybridization assay and HC-Ⅱwas 92.4%.The most common HR-HPV subtypes among patients with CIN2 or above were HPV16,18,52,58, 31.The positive rate of HPV 16 increased rapidly with the severity of the disease.The most common HPV types were 16,52,11,18,58 in CINⅡ,group,16,58,18,52,31 in CINⅢgroupand 16,18,31,52,58 in ICC group.2.The growth of Caski and H8 was inhibited by different concentration of Baofukang suppository.After Baofukang Suppository treatment,Caski and H8 cell line showed decreased levels of HPV16 E6E7 mRNA than that in the control group.The growth of CaSki,SiHa and H8 was inhibited by different concentration of curcuma aramatica oil.SiHa cells were accumulated in G2,S phase(P＜0.01),while CaSki and H8 cells were accumulated in S phase(P＜0.01).The apoptosis rate in CaSki,SiHa and H8 cells was higher than that in control group(P＜0.01, P＜0.01,P＜0.05).CaSki,SiHa and H8 cell showed decreased level of HPV16 E6E7 mRNA than that in the control group after curcuma aramatica oil treatment(P＜0.01).Conclusions1.The positive rate of HR-HPV increases with the severity of the cervical lesions.HR-HPV testing is useful in triage of ASC-US,while its triage role for ASC-H need further study.The HR-HPV load is significantly associated with the presence of CINs and ICC.HPV viral load does not increase with severity of cervical neoplasia.The 5 most common subtypes in patients≥CIN2 or above are HPV16,18,52,58,31.2.The results suggest that Baofukang Suppository and curcuma aramatica oil may execute its antiHPV activity primarily by decreasing the expression of HPV 16E6E7 in cervical cancer cell line and immortalized cervical epithelial cell.