| Objective: Ovarian cancers are the most common type of cancer inwommen, leading the mortality rate among gynecological cancers. However,there was no sufficiently sensitive tests to diagnosis ovarian cancer,70%of thepatients are diagnosed at an advanced stage,and the five year survival rate isonly30%-40%.85%-90%of malignant ovarian tumors are epithelial ovariancancer in origin,therefore it is important to detect EOC at an early stage. Atpresent CA125is the most frequently used biomarker for EOC detection,but itis only elevated in approximatly50%of stage Ⅰ EOC and70-90%of advancedcases. In the earliest publications,it was appreciated that CA125could beelevated in several benign diseases and in patients with nonovarianmalignancy,including cancers of the endometrium,lung,breast, pancreas,andcolon. Because of the low sensitivity and false positive of CA125,it isimportant to identify new serum biomarkers that alone or in combination withCA125,could improve early EOC diagnosis. Mitochondrial DNA play animportant role in inherit outside of the nuclear,because of lack of protectivehistones and limited capacity for DNA repair,somatis mtDNA mutations occurin a wide varity of degenerative diseases and cancers. Somatic mtDNAmutations and polymorphisms with a higest prevalence in the D-LOOP controlregion,SNPs in the mtDNA D-LOOP were found to be risk markers forepithelial ovarian cancer,which have been studyed,the aim of this study was toevaluate SNPs of mtDNA D-LOOP in combination with CA125in EOCdiagnosis at an early stage.Methods:1samples collection and patients follow up:①case group:93patients who had presented for treatment of epithelial ovarian cancer to the SecondHospital of Hebei Medical University,between2005and2009,all the patientswere not given any treatment before surgery.②controls: mtDNAD-LOOP of93healthy females.2Total DNA was extracted using a DNA blood extraction kit. The purityand concentration of DNA were analyzed spectrometrically.3PCR amplification of mtDNA D-LOOP.4After amplification,4μl of PCR product was electrophoresed on a1%agarose gel to confirm specific amplify-cation.5Cycle sequencing was carried out with the Dye Terminator CycleSequencing Ready Reaction Kit and the products were then separated on theABIPRISM Genetic Analyzer3100(Applied Biosystem). Mutations andpolymorphism were confirmed by repeated analysis.6statistical analysis:We used the χ2test to analyze the markers in thedetection of EOC at an early stage.The level of P≤0.05was consideredsignificant. And we also use a further analysis of significant Chi-Squrae Testfor Multiple Sample Rate comparison,a P-value<0.0125was considered toindicate statistical significance.Results:1There were no statistical difference of age,Gravidity,Parity betweenepithelial ovarian cancer patients and healthy controls,but CA125was signif-icantly higer in EOC patients compared with healthy controls(P≤0.05).2Among all the patients,SNPs were detected in187sites within the982-bp at the mitochondria D-loop region from the blood samples of thehealthy controls and epithelial ovarian cancer patients,11mutations and176polymorphisms were identified.And the mutation rate is64.5(60/93). Whenindividual SNPs were analyzed comparing epithelial ovarian cancer patients tocontrols,a statistically significant increase in SNP frequency for the73A/G(relative risk,123.760;95%CI,28.338-540.495;P=0.000),207G/A (relativerisk,4.875;95%CI,1.024-23.213;P=0.030)and523C/del (relative risk,1.982;95%CI,1.089-3.605;P=0.024),which indicated that the patients who carry these alleles were susceptible to epithelial ovarian cancer. Additional SNPs(254T/G,259A/G,275G/A,366G/A,411C/G,414T/G,418C/G,441C/A,476C/A,524C/del,530C/T)were significantly associated with the tendency toward theincreased risk for epithelial ovarian cancer. In contrast,the allele249A/del(relative risk,2.406;95%CI,1.207-4.797;P=0.011)and263A/G (relative risk,3.616;95%CI,1.133-11.541;P=0.022)was correlated with the reduced risk ofepithelial ovarian cancer.3According to the Chi-Squrae Test for Multiple Sample Rate comparison,the sites of73A/G,523C/del,366G/A,411C/G524C/del,which showed CA125and different SNPs sites are a potential role for early stage detection of EOC.Conclusion:1In conclusion,SNPs in the mtDNA D-LOOP were found to be riskmarkers for epithelial ovarian cancer.2In conclusion,This study confirms the diagnostic role of CA125and73A/G,523C/del,366G/A,411C/G,524C/del,and for the first time showed thatCA125and73A/G,523C/del,366G/A,411C/G and524C/del are a potentialrole for early stage detection of EOC. |
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