INVESTIGATION OF MELATONIN RECEPTOR GENE AND PROTEIN EXPRESSION IN HUMAN HUT78 CELL LINE
- VernacularTitle:人HUT78细胞褪黑素受体的基因与蛋白表达的研究
- Author:
Xiangfang CHEN
;
Zhimin LIU
;
Ying ZHAO
- Publication Type:Journal Article
- Keywords:
HUT78 cell line;
receptors, melatonin;
mt 1;
MT 2;
immunohistochemistry
- From:
Medical Journal of Chinese People's Liberation Army
1982;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the melatonin receptor(MR) gene and protein expression in human HUT78 cell line, total RNA of HUT78 cells was isolated by single-step method of acid guanidinium-thiocyanate-phenol-chloform, and mt 1 and MT 2 mRNA were determined by reverse transcription-polymerase chain reaction(RT-PCR). The Envision method was applied to immunohistochemistry to identify and localize mt 1 and MT 2 protein. The mt 1 cDNA fragment of the expected size of 370bp was determined, but the MT 2 cDNA fragment of the expected size of 320bp was not determinable by RT-PCR. Sequencing result indicated that the positive product coincided with the cDNA of human mt 1. The mt 1 protein was observed by immunohistochemistry. These buffy positive granules were scattered with some areas stronger than the others, and were primarily located in cytoplasm and membrane, with rare location in nucleus; meanwhile the MT 2 protein was not observed. These results demonstrated the mt 1 mRNA and protein expression in HUT78 cell line. It is indicated that melatonin has directly immune-regulative effects on T lymphocyte, The changes of MR in physiological and pathological stages need to be investigated.
