| Background and objectiveCervical cancer was one maligant tumor of female. For sexual transmitted diseases and HPV infection, the patients with cervical cancer were becoming younger and younger. For the wide-ranging general survey used in cervical cancer, the rate of earlier diagnose of cervical cancer was improved. And the patients' survival rate have been increased.As the literature reported the detection rate of HPV in cervical cancer was over 90 percentage. HPV is a double-stranded circular structure DNA virus and it is made up of about 8000 base pairs. The major cause of cervical cancer was the original cancer protein encoded by E6 gene and E7 gene of high risk HPV. They are small protein molecules, which contain 158 and 98 amino acids. They have zinc finger structure, which related to the malignant transformation of cells closely. Cells in vitro, which expressed original cancer protein of E6 gene and E7 gene, would reach eternal life. And the eternal life would be a prelude of tumor. Transgenic study also founded that in high-risk HPV E6 gene and E7 gene could directly lead to tumor. For the reasons above, some scholars have already successfully developed the HPV vaccine. With clinical application of HPV vaccine, the incidence of cervical cancer would be significantly decreased. Gene therapy would be a trend in clinical in the future. For the research about specific gene of cervical cancer was few, the progress of gene therapy in cervical cancer was inhibited. WAPL gene was found in Drosophila. And in human the homology sequence was HWAPL gene. The specific role of HWAPL gene may be keeping the stability of chromosome. Oikawa and so on found that compared to ovarian cancer, breast cancer, lung cancer, kidney cancer, colon cancer and the corresponding normal tissues the expression of HWAPL mRNA in cervical cancer increased more obviously. They also found that compared to other cell lines, HWAPL gene in cervical cancer cell lines expressed a higher level. So they guessed that HWAPL gene may be a cervical cancer-specific gene and this promoted the research progress of gene therapy in cervical cancer.In this test, HPV would be detected and typed from paraffin-embedded specimens of cervical cancer, then provided a theoretical basis for studying HPV. At the same time, immunohistochemical results of HWAPL protein would be compined to find out the relationship between HPV and HWAPL gene expression. This study will provide a theoretical basis for the prevention and gene therapy of cervical cancer in the future.Materials and methods75 cases of cervical paraffin-embedded specimens were collected in Zhengzhou University 2nd affiliated hospital from February 2004 to February 2006. The age of 30 cases of cervical squamous cell carcinoma was from 29 years old to 57 years old and the average age was 43 years old. In 45 cases of cervical intraepthelial neoplasia , there were 15 cases of CIN I,15 cases of CIN II and 15 cases of CINIII. The age of 45 cases of CIN was from 24 years old to 50 years old and the average age was 37 years old. All women hadn't been treated with chemotherapy or radiotherapy.PCR and gene chip technology were used to detect and type HPV from paraffin-embedded cervical specimens. According to typing results and immunohistochemical results of HWAPL protein, we would find the relationship between HPV with HWAPL gene expression. SPSS10.0 was used to analyze the datum.Results (1) After amplifying with PCR, the results showed that there were 28 cases of HPV DNA positive in 30 cases of cervical cancer and the positive rate was 93.3%. In 15 cases of CIN III, there were 14 cases of HPV DNA positive and the positive rate was 93.3%. In 15 cases of CIN II, there were 14 cases of HPV DNA positive and the positive rate was 93.3%. In 15 cases of CIN I , there were 12 cases of HPV DNA positive and the positive rate was 80%. Detected by gene chip, the results showed that in 30 cases of cervical cancer there were 27 cases of HPV16/18 positive and 1 case of HPV6 positive. The positive rate was 90% and 6.7% respectively. In 15 cases of CIN III, there were 10 cases of HPV16 /18 positive, 1 case of HPV58 positive and 3 cases of HPV6/11 positive. The positive rate was 66.7%, 6.7% and 20% respectively. In 15 cases of CIN II, there were 11 cases of HPV6/11 positive and 3 cases of HPV16/18 positive. The positive rate was 73.3% and 20% respectively. In 15 cases of CIN I , there were 10 cases of HPV6/11 positive and 2 cases of HPV 16/18 positive. The positive rate was 66.7% and 13.3% respectively. In cervical cancer and CIN III the detection rate of high-risk HPV was significant higher than in CIN II and CIN I (x~2 = 32.248, P=0.000).(2) The immunohistochemical results of HWAPL protein showed that in 30 cases of cervical cancer there were 5 cases with weak positive staining intensity (+), 8 cases with middle positive staining intensity (++) and 17 cases with strong positive staining intensity(+++). In 15 cases of CIN III, there were 4 cases with +, 4 cases with ++ and 7 cases with +++. In 15 cases of CIN II, there were 6 cases with +, 5 cases with ++ and 4 cases with +++. In 15 cases of CIN I , there were 7 cases with +, 6 cases with ++ and 2 cases with +++. There were significant difference between the expression of HWAPL protein in CIN III and cervical with the expression of HWAPL protein in CIN II and CIN I (x~2 = 8.928, P=0.012).(3) There were 7 cases of HPV negative specimens, in which 4 cases with HWAPL protein +, 2 cases with ++ and 1 case with +++. There were 26 cases of low risk HPV positive specimens, in which 12 cases with HWAPL protein +, 5 cases with ++ and 9 cases with +++. There were 42 cases of high risk HPV positive specimens, in which 6 cases with HWAPL protein +, 12 cases with ++ and 24 cases with +++. Specimens with high-risk HPV positive expressed HWAPL protein significant higher than specimens without high risk HPV positive (x~2=14.844, P=0.005). Correlation analysis showed that high risk HPV and the expression of HWAPL gene correlated (r =0.440, P= 0.000).Conclusions(1) 90% HPV DNA could be detected from Paraffin-embedded specimens of cervical cancer. And these specimens could be used as cervical HPV infection research.(2) For automation throughput high accuracy and specificity advantages, gene chip technology could be used as an important method for clinical HPV testing and classification.(3) In cervical cancer HWAPL gene expression was high. High risk HPV infection may increased the replication and expression of HWAPL gene. The abnormal expression of HWAPL gene may play an important role in the pathogenesis of cervical cancer.
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