重组截短型TGF-β前体潜在相关肽的原核表达及其对HSC-T6细胞增殖的影响

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Abstract ObjectiveTo preliminarily study the influence of recombinant protein of latency-associated peptide(LAP)of recombinant truncated TGF-β precursor on proliferation of hepatic stellate cells　(HSC)-T6.MethodsThe truncated LAP fragment was amplified by polymerase chain reaction(PCR)and the prokaryotic expression vector was constructed.After double enzyme digestion and DNA sequencing,the protein expression was induced by IPTG at 0.5 mmol/L at 37°C for 4 h.The expressed protein was detected by SDS-PAGE and Western blot,after that the recombinant protein was by nickel agarose affinity chromatography for purification.The recombinant protein was applied to rat hepatic stellate cell-T6　(HSC-T6),and the influence of different concentrations of recombinant protein on cell proliferation was detected by MTT assay.ResultsThe prokaryotic expression vector was successfully constructed and the recombinant protein was expressed and purified successfully.The proliferation of HSC-T6 cells decreased with the increase of the recombinant protein concentration.ConclusionThe recombinant protein inhibits the proliferation of HSC-T6 cells,and lays the foundation for the subsequent study on the interference of LAP fragments on the activation of TGF-β and the influence on organ fibrosis.

Key words： Latency-associated peptide of recombinant truncated TGF-&beta precursor;Prokaryotic expression;Protein purification;Hepatic stellate cells-T6;Proliferation

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	Articles by authors
	SHIYong-ping
	ZHENGJun-ya
	LIUZeng-rui
	LILu-xin
	WANGJing
	LILing-yu
	CAOYa-nan
	CHUYan-hui

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SHIYong-ping,ZHENGJun-ya,LIUZeng-rui, et al. Prokaryotic expression of latency-associated peptide of recombinant truncated TGF-β precursor and its influence on proliferation of HSC-T6 cell[J]. 中国当代医药, 2018, 25(17): 4-7.

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http://dangdaiyiyao.com/EN/ OR http://dangdaiyiyao.com/EN/Y2018/V25/I17/4

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