人膀胱移行细胞癌T24细胞中CK20mRNA表达检测方法的建立

Established a method of detecting the expression of CK20 mRNA in T -24 cell line of bladder cancer

目的建立巢式RT—PCR方法检测人膀胱移行细胞癌T24细胞中CK20。RNA表达进行分析的实验方法。方法培养人膀胱移行细胞癌T24细胞，利用普通RT—PCR及巢式RT—PCR方法分别检测其CK20mRNA的表达。结果1．5％琼脂糖凝胶电泳发现普通RT—PCR及巢式RT—PCR结果均出现与预期结果一致的阳性条带。结论巢式RT—PCR方法检测人膀胱移行细胞癌T24细胞中有CK20mRNA的表达。为进一步利用巢式RT—PCR方法研究临床膀胱移行细胞癌患者尿脱落细胞及肿瘤组织中CK20mRNA的表达、提高检测敏感性及可靠性打下基础。为膀胱癌早期诊断和术后监测提供一种新的有效手段。

Objectives To establish an experimental method detecting the expression of CK20 mRNA in T -24 cell line of bladder cancer with nested reverse transcription -polymerase chain reaction （nested RT- PCR ）. Methods T - 24 cell line of bladder cancer were cultivated. CK20 mRNA was detecied by RT - PCR and nested RT- PCR. Results CK20 amplification band （370bp and 210bp） was obtained with mRNA extracted from T - 24 cell line of bladder cancer. Conclusions Nested RT - PCR is a method of detecting the expression of CK20 mRNA in T -24 cell line of bladder cancer. It may provide an experimental method for studying the expression of CK20 mRNA in urine exfoliated cells of transitional cell carcinoma of the bladder （TCC） and their tumor tissues in that we can improve the sensitivity and possibility, which can provide a new effective way in early diagnosis and fol- low - up of TCC.