A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment

QIAN Yujun; QIN Chunxia; SUN Lili; DING Huamin; LI Tiejun

doi:10.3969/j.issn.1006-0111.2019.01.007

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Objective To establish a high-throughput in-vitro screening cell model for anti-atherosclerosis leading compounds. Methods Hypoxia response element (HRE) was cloned into a luciferase reporter vector, pGL3-Enhancer, to construct pGL3-HIF-1α-HRE. The THP-1 human monocyte cell line was infected with the pGL3-HIF-1α-HRE and a stable cell line, THP-1-HIF-1α-HRE, was screened. Results Real-time PCR assay showed that HIF-1α expression and luciferase activity in THP-1-HIF-1α-HRE cells was effectively upregulated by hypoxia. The increase of HIF-1α expression and luciferase activity induced by hypoxia was significantly inhibited by lovastatin or curcumin. Conclusion THP1-HIF-1α-HRE, an in-vitro cell model for high-throughput screening lead compounds for anti-atherosclerosis (AS) was successfully established.

A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment

Department of Pharmacy, Punan Hospital of Pudong New Area, Shanghai 200125, China

Received Date: 2018-06-11

Rev Recd Date: 2018-09-25

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Abstract: Objective To establish a high-throughput in-vitro screening cell model for anti-atherosclerosis leading compounds. Methods Hypoxia response element (HRE) was cloned into a luciferase reporter vector, pGL3-Enhancer, to construct pGL3-HIF-1α-HRE. The THP-1 human monocyte cell line was infected with the pGL3-HIF-1α-HRE and a stable cell line, THP-1-HIF-1α-HRE, was screened. Results Real-time PCR assay showed that HIF-1α expression and luciferase activity in THP-1-HIF-1α-HRE cells was effectively upregulated by hypoxia. The increase of HIF-1α expression and luciferase activity induced by hypoxia was significantly inhibited by lovastatin or curcumin. Conclusion THP1-HIF-1α-HRE, an in-vitro cell model for high-throughput screening lead compounds for anti-atherosclerosis (AS) was successfully established.

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A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment

doi: 10.3969/j.issn.1006-0111.2019.01.007

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通讯作者: 陈斌, [email protected]

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