原位杂交结合免疫组化研究Ki-67基因的表达和调控

A study on the expression and regulation of Ki-67 gene by in sift hybridization combining with immuno-histochemistry

为研究Ｋｉ－６７基因的表达和调控，我们应用原位杂交和免疫组化研究Ｋｉ－６７抗原的ｍＲＮＡ转录和蛋白表达的相关性。采用常规福尔马林固定和石蜡包埋的人类扁桃体，地高辛标记的ｃＲＮＡ探针进行ｍＲＮＡ原位杂交，杂交信号显示方法为直接免疫磷酸复和物法（ｄｉｒｅｃｔｉｍｍｕｎｏａｋａｌｉｎｅｐｈｏｓｐｈａｔａｓｅ，ＩＡＰ）。结果发现：地高辛标记的Ｋｉ－６７ｃＲＮＡ探针首次成功地检测到常规处理的扁桃体石蜡切片中的Ｋｉ－６７ｍＲＮＡ，Ｋｉ－６７ｍＮＡ的转录和其蛋白表达存在着相关性。结果说明：本实验中建立的方法有助于深入地分析Ｋｉ－６７基因表达和调控。

In situ hybridization and immunohis- tochemistry were used to investigate the correlation be- tween the mRNA transcription of Ki-67 antigen and its expression of protein. The routinely formalin-fixed and paraffin-embedded human tonsil tissues were treated to make in situ hybridization with digoxigcnin (DIG ) -lab- clcd Ki-67 antigen cDNA probes. The DIG-labeled cR- NA probes were detected by means ef a direct im- munoalkaline phosphatase (IAP ) procedure using 4-nitro blue tetrazolium chloride and 5-bromo-4-chloro-3-in- doylphosphatase ( NBT/BCIP ) as the substrates, The mRNA of the Ki-67 antigen in the tonsil tissues was successfully detected , which implied the existing correla- tion between the transcription of the Ki-6 7 mRNA and its protein expression. The method presented here may be valuable for further analysis of the Ki- 67 antigengene expression and regulation.