The MRNA Expression Level And Siganificance Of Kinectin Alternative Splicing Regions Of ORF In Normal Tissues And HCC Tissues

OBJECTIVE: To investigate the mRNA expression of hepatocellular carcinoma related antigen kinectin alternative splicing regions of open reading framework(ORF)in normal tissues,To investigate the mRNA expression of kinectin alternative splicing regions in HCC tissues , corresponding adjacent liver tissues and normal hepatic tissues.,To discuss the correlation of the expression of kinectin alternative splicing regions with HCC.METHOD:①The mRNA expression of kinectin predictive six altenative splicing regions in ORF were detected with reverse transcription polymerase chain reaction (RT-PCR) in 16 kinds of human normal tissues.②The mRNA expression of kinectin V2 altenative splicing region was detected in 46 cases of HCC tissues and corresponding adjacent liver tissues,9 cases of normal hepatic tissues,9 cases of retinoblastoma tissues,7 cases of normal retinal tissues,9 cases of glioma tissues and 9 cases of normal brain tissues. The mRNA expression of kinectin V4 altenative splicing region was detected in 10 cases of HCC tissue and corresponding adjacent liver tissue ,9 cases of normal hepatic tissues;The mRNA expression of kinectin V1 altenative splicing region was detected in 9 cases of glioma tissues and 9 cases of normal brain tissues . Correlation between the ratio of kinectin V2L/V2S (L was referring to large product fragment containing V2 splicing region,S was referring to short product fragment not containing V2 splicing region)in HCC tissue and clinical data was evaluated.Genes of positive PCR results were sequenced randomly.RESULT:①Of 16 human normal tissues, only three alternative splicing regions V1,V2 and V4 were detectable. The alternative splicing region V1 was existing restricted in brain tissue,The alternative splicing region V2 and V4 were existing widely in all normal tissues.②It was intresting that the ratio of V2L/V2S were 0.706±0.204 , 0.543±0.162 and 0.553±0.231 in HCC tissues ,corresponding adjacent tissues and normal hepatic tissues respectively. There were significant statistical difference. But the ratio of V2L/V2S were 1.836±0.285,1.429±0.513,1.047±0.409 and 1.273±0.624 in retinoblastoma tissues, normal retinal tissues, glioma tissues and normal brain tissues ,respectively. There were no statistical difference between retinoblastoma tissues and normal retinal tissues, glioma tissues and normal brain tissues.The ratio of V4L/V4S were 0.966±0.195 , 1.015±0.154 and 0.873±0.162 in HCC tissues ,corresponding adjacent tissues and normal hepatic tissues,There were no statistical difference. and the ratio of V1L / V1S were 1.833±0.345,1.454±0.208 in glioma tissues and normal brain tissues respectively,There were no statistical difference. In HCC tissues, the ratio of V2L/V2S was related with the size of HCC,clinical TNM staging. DNA sequencing confirmed that the PCR products were true target cDNA.CONCLUSION: In human normal tissues, there are existence of three alternative splicing regions such as V1,V2 and V4, the alternative splicing region V1 is existing restricted in brain tissue,the alternative splicing region V2 and V4 are existing widely in normal tissues. In HCC tissues,the increased ratio level of the kinectin V2L/V2S may correlate with the recurrence and metastasis of HCC.