Studies On Candidate Upregulated Genes As Markers For Early Prognosis Of Cervical Cancer

Objective:Early detection is the key to the treatment of cervical cancer,and screening of tumor molecular markers is an effective way to establish the method of early diagnosis of tumors.In this study,we screened candidate upregulated genes in Uyghur women with cervical carcinoma by whole human genome microarray approach,and further detected the specificity and sensitivity of candidate genes for the evaluation of the reliability and accuracy of transcriptomics,and the establishment of markers for early prognosis of cervical cancer.Methods:(1)We collected 72 cases of fresh tissue specimens from Uyghur women with cercical lesions,including 25 cases of cervical squamous cell carcinoma(CSCC),22 cases of cervical intraepithelial neoplasia(CIN)and 25 cases of cervicitis or normal cervix(NC);(2)20 cases of high quality tissue specimens(7 cases of CSCC,6 cases of CIN III and 7 cases of NC)were selected for the screening of candidate up-regulated genes specific to cervical carcinoma and its precursors by extraction tissue RNA and subsequent analysis with human genome-wide expression profile.(3)3 candidate up-regulated genes were selected for the design of mRNA sequence specific primers and subsequent detection of candidate gene transcription in 72 cases of tissue RNA by semi-quantitative and fluorescence quantitative RT-PCR,and to assess the specificity and sensitivity of candidate gene detection for the early diagnosis of cervical cancer,and evaluate reliability and accuracy of the results of microarray analysis.Results:(1)By analysis of 20 cases of cervical tissue RNA by human genome expression profile chip,and setting up at least two times of difference in gene expression as quantitative standard for comparative analysis,we identified 394 or 516 genes up-regulated in CSCC tissues,respectively,compared to CIN? or NC,whereas no up-regulated genes were found for CIN ? in relation to NC.(2)We selected 3 candidate genes up-regulated in cervical cancer including SIX1,DTL and APOBEC3B for the screening analysis of tissue RNAs by semi-quantitative RT-PCR,and the results have shown that the expression of all genes above was up-regulated with the development of cervical lesions,with significant differences in up-regulation of 3 genes at transcriptional level,in CSCC compared to CIN?-? or NC(P<0.05),whereas no statistical difference was found in CIN?-? relation to NC(P>0.05).(3)Further analysis of 3 candidate gene expression by fluorescent quantitiative RT-PCR showed that the mRNA expression of SIX1 and DTL genes was the highest in CSCC(0.891±0.517 and 0.026±0.021),decreased in CIN?-?(0.53410.290 and 0.015±0.007)and the lowest one in NC(0.035±0.032 and 0.003±0.003),and pair wise comparison was statistically significant(P<0.05)in any other group except for DTL which has no difference between CSCC and CIN ?-?,but no statistical difference was for APOBEC3B.The statistical analysis has shown that the sensitivity of SIX 1 and DTL detection had a relatively high sensitivity for the diagnosis of cervical cancer,but with relatively low specificity and accuracy(95.7%,42.6%and 54%v.s.96.0%,21.7%and 38.9%)respectively.Analysis of the correlation of two genes showed a moderate correlation(r=0,490,P=0.000).Conclusion:The development of cervical cancer in Uighur women is associated with the up-regulation of multiple genes.The changes in SIX1 and DTL expression may be markers of cervical cancer,and the detection of DTL may have a relatively high impact in the prognosis of cervical cancer,may also be a proof for the feasibility and reliability of data resulted from the transcriptomics screening analysis.