| Purposes:1Taking clinical and pathological findings as a gold standard, the thesis analyses and compares the prediction values of Thinprep Cytology, HPV infection, hTERC, and c-myc gene amplification in cervical lesions through the test of hTERC gene, HPV subtypes, c-myc gene and classification of Thinprep Cytology in exfoliated cells of cervical epithelium.2The thesis is designed to seek a more accurate test method to diagnose and predict cervical disease through the analysis and comparison of examination results of the c-myc gene, hTERC gene, HPV subtypes and ThinPrep Cytological Test.3The thesis discusses the values of hTERC, c-myc gene, HPV subtypes as well as TCT in the follow-up after operation in CIN patients who had experienced conization or hysterectomy treatment.Methods We chose1000women, aged25to64years old, who have been married and have sexual life history from those who doctored in department of gynecology of the First Affiliated Hospital of Chongqing Medical University in January to September2011. The selected patients were brushed the cervical exfoliated cell for liquid-based cytology, the residual liquid at the same time as to detect c-myc and hTERC gene expression by fluorescence in situ hybridization, by surface plasmon resonance to detect HPV types, positive patients with any of the indicators as liquid-based cytology, HPV subtypes, hTERC gene and c-myc gene underwent colposcopy and biopsy.After screening, at last117patients required colposcopy and biopsy, but2of them gave up. Therefore, only115patients had pathological results. The patients with Normal/inflammation and CINI directly followed up without treatment; The patients with CINII, CINIII, SCC were taken to surgery and follow-up after6month. During the process of follow-up,26who are mainly inflammation and hysterectomy patients missed the follow-up. There were89taking referral. They were made c-myc, hTERC gene, HPV subtype and TCT test of exfoliated cervical cells in6months. When necessary, they were taken colpscope and biopsy.Results:1The positive rates of CINI, CINII, CINIII, SCC patients in TCT test are respectively87.5%,96.4%,100%,100%. With the hypertension of pathological level, the TCT positive rates are increasing. But for different levels of CIN screening, there is no obvious superiority (P=0.061).2with the increasing severity of the lesion histopathology, c-myc gene amplification rates are increasing, and c-myc gene positive amplification rate are also increasing. In the inflammatory/normal, CINI, CINII, CINIII, SCC group, the positive rates of c-myc gene amplification are as follows:41.67%,18.75%,39.29%,54.90%,87.50%. It shows that the statistics of the c-myc gene amplification positive rate of different pathological levels is significant(P<0.05).3The levels of pathological detection are inflammation\normal, CINI, CINII, CINIII, SCC. The positive rates are respectively:33.33%,25%,50%,68.63%,87.50%. As the lesion severity increases, the positive rate of hTERC gene amplification is also increasing.4As the pathology level increases, the positive expression rates of HPV gene amplification of inflammation, CINI, CINII, CINIII, SCC patients are respectively:75%,68.75%,100%,88.24%,87.50%. The results show that the positive expression of HPV in different pathological levels, the difference was statistically significant.5the test sensitivities of TCT, HPV, c-myc gene, hTERC gene in cervical leision are respectively86.21%,92%,52.9%,64.40%; specificities are70%,26.70%,70.00%,70.00%. The sensitivities of TCT+c-myc, TCT+hTERC, HPV+c-myc, HPV+hTERC, c-myc+hTERC are93.50%, 95.09%,96.23%,97.15%,83.28%; specificities:91.00%,91.00%,78.01%,78.01%,91.00%. The sensitivities of TCT+HPV+c-myc, TCT+HPV+hTERC, TCT+HPV+c-myc+hTERC are99.48%,99.61%,99.81%; specificities:93.40%,93.40%,98.20%. The HTERC, c-myc gene test sensitivity and specificity are not high, but the specificity and sensitivity increase to a certain extent when combining HPV and TCT. Besides, Compared with c-myc gene, hTERC gene are independently tested and TCT+HPV detection in sensitivity and specificity to show greater advantages; four joint sensitivity and specificity were as high as99.81%and98.02%, but at the same time detection of two genes increased the economic burden of patients.6Of the69CIN patients with conization,3failed in follow-up, and66took referral in6months. All the66patients’ positive rate of TCT, HPV, c-myc gene, hTERC gene reduced obviously comparing the results of before and after treatment.Conclusions:1The amplification of c-myc gene and hTERc gene may be the early event during the formation of cervical cancer. C-myc gene and hTERc gene, which play a very important role in the process of cervical cancer, can be considered as the molecular marker of CIN screening. The amplification of two genes has an important reference value on predicting CIN recurrence or progression. They also can be used as a follow-up method after CIN treatment.2HPV infection is closely related to the cervical intraepithelial neoplasia. Especially, HPV high-risk patients have a higher rate CIN incidence. The detection of high-risk HPV has a significant meaning in the examination of cervical lesions. Because of the higher rate of HPV infection and the lower specificity, the combination between the detection of high-risk HPV and the cervical cytology test can obliviously increase the detection rate of cervical lesions. The method is also considered as the major joint screening of CIN.3The HPV infection and the rate of amplification of C-myc gene and hTERc gene are positively correlated in the beginning of cervical lesions, the amplification of C-myc gene and hTERc gene and the HPV infection play a very important role in the process of cervical lesions and cervical carcinoma. Their abnormal expressions in cervical lesions are associated. The combination between the detection of C-myc gene, hTERc gene and the commonly used detection of TCT+HPV has a higher sensitivity and specificity. Thus TCT+HPV+hTERC or TCT+HPV+c-myc are thought as the best combinations.4FISH technology has the advantages of being objective, repeatable and producing no injuries. Moreover, it screens cervix lesions by taking advantage of exfoliated cervical cells, detects the c-myc and hTERC gene. It makes up the low sensibility of TCT and low specificity of HPV and can be taken as an effective detection method of the "three-step".5This research utilizes SPR to make HPV DNA detection in exfoliated cervical cells. Distinctions in HPV positive expressions in different levels of pathology have statistics significance, but they are not positively correlated with levels of histopathology.lt may be related to insufficient clinical cases and detection methods which needs further research by the means of SPR technology. |
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