摘要
目的探索TSGF和hMAM表达指标对乳腺癌细胞外周血微转移诊断的应用价值。方法分离受检者外周血单个核细胞,TRIZOL液提取总RNA,作RT-PCR并用-βactin监控;确认的cDNA先作普通PCR扩增,再作荧光定量PCR扩增。血清作TSGF检测。结果64例标本有63例确认转录为cDNA,其中乳腺癌43例、良性乳腺肿瘤和正常对照各10例。经普通PCR后再作荧光定量PCR,乳腺癌hMAM表达阳性率为53.3%、良性乳腺肿瘤和正常对照均无阳性,乳腺癌组与良性乳腺肿瘤或正常对照组间hMAM表达有显著差异;淋巴结转移组与未转移组,hMAM表达有显著差异。TSGF阳性率为1.6%,乳腺癌组与良性乳腺肿瘤组及正常对照组间无显著差异。结论二次PCR的灵敏度可成功检出外周血hMAM的表达,而单次PCR法的灵敏度不够;外周血hMAM表达指标可用于监测乳腺癌细胞微转移,但未见TSGF指标对乳腺癌微转移的监测价值。
Objective To evaluate the application value of tumor supplied group of factors（TSGF） and human mammaglobin （hMAM） expression in peripheral blood for detecting micrometastasis of breast cancer. Methods mononuclear cells from patient＇s peripheral blood were isolated. Total RNA was extract by using TRIZOL, RT PCR was then performed. β-actin gene amplification was used as control, Both PCR and Real-time PCR were applied after cDNA amplification was achieved in dais study. TSGF was detected from blood serum. Results 63 of 64 samples had cDNA amplification, Among them, 43 cases were breast cancer, and 10 of them were breast benign lesions and normal controls, By using real time PCR as second round amplification in a nested PCR, hMAM expression was detected in 53.3% of breast cancer patients tested, showing statistic difference（ P 〈 0.05） compared with the breast benign lesions and normal controls. No any positive results were found when only single round real time PCR was applied, Higher hMAM expression was detected in patient group with lympho-nude metastasis, showing statistic difference（ P 〈 0.01 ） compared with patient without lympho-nude metastasis, Positive rate of TSGF was 1,6% in breast cancer patients, which has no significant difference（ P 〉 0.05） compared with the breast benign lesions and normal controls, Conclusion Human manmmaglobin（hMAM） expression in the peripheral blood can be an indicator of the micrometastasis of breast cancer cells, Nested PCR with combination of 1st normal PCR and 2nd real time PCR has improved sensitivity in hMAM gene amplification, TSGF has not been found any value for detecting micrometastasis of breast cancer cell.
出处
《中国实验诊断学》
2005年第4期536-538,共3页
Chinese Journal of Laboratory Diagnosis
基金
浙江省医学科学研究基金(2004A086)
关键词
乳腺癌
人乳房珠蛋白
RT-PCR
TSGF
外周血
breast cancer
Human mammaglobin（hMAM）
RT PCR
tumor supplied group of faetors（TSGF）
peripheral blood
