荧光定量PCR技术检测乳腺癌ck19 mRNA表达过量的意义

Clinical application of fluorescent quantification polymerase chain reaction to detect over expression of CK19 in breast cancer

目的 探讨荧光定量PCR(FQ PCR)检测细胞角蛋白 19(ck19)在乳腺癌诊断中的应用。方法 采用FQ PCR法 ,并以GAPDH为内对照测定 15例健康女性体检者、2 5例良性乳腺疾病患者和 76例乳腺癌患者的外周血中ck19mRNA的表达量。结果 正常对照组和良性乳腺疾病组ck19和ck19/GAPDH无显著性差异 ,乳腺癌组均显著高于前 2组 ,GAPDH在 3组间均无显著性差异。若以ck19基因表达量和ck19/GAPDH高于正常对照组 x +2s为阳性 ,76例乳腺癌患者的阳性数为 4 1,阳性率5 3.9% ,良性乳腺疾病组阳性数为 0。 76例乳腺癌中 2 7例患者从手术前到第 3次检测 ,ck19和ck19/GAPDH均显下降趋势 ,但第 1次和第 2次与手术前比较 ,无显著性差异 ,而第 3次与手术前比较 ,ck19和ck19/GAPDH均显著低于手术前 ,存在显著性差异。结论 FQ PCR技术是高度灵敏、高度特异的快速定量检测ck19基因的方法 ,可有效监测乳腺癌的诊断。

Objective To evaluate the clinical application of fluorescent quantification polymerase chain reaction (FQ PCR) to detect cytokeratin 19(CK19) in the diagnosis of breast cancer.Methods Clinical samples of 15 health women, 25 patients with benign breast disease and 76 patients with malign breast disease were detected by FQ PCR.GAPDH was used as internal control.Results There were no significant difference in ck19 and ck19/GAPDH between normal controls and benign breast disease group.ck19 and ck19/GAPDH in breast cancer group were higher than those in the other groups.There was significant difference between them.There were no difference in GAPDH among three groups.Using the standard that the expression of ck19 and ck19/GAPDH was higher than ±2 s of normal controls, the positive ratio of breast cancer and benign disease were 53.9 and 0 percent respectively.ck19 and ck19/GAPDH in 27 breast cancer patients were declining after chemotherapy.There were no difference between pre operation and first and second chemotherapy.There was significant difference between pre operation and third chemotherapy.The latter was lower than the former.Conclusions FQ PCR can detect ck19 with high specificity and sensitivity.It also gives objective evidence to the diagnosis, therapy and prognosis of breast cancer