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缺氧環(huán)境下HSulf-1,HIF-1α在HaCaT細胞中的表達及HSulf-1過表達載體的構(gòu)建

發(fā)布時間:2019-03-14 14:26
【摘要】:目的探討HSulf-1,HIF-1α在缺氧時人角質(zhì)形成細胞株HaCaT細胞中的表達并構(gòu)建HSulf-1基因過表達載體。方法 Western blot法檢測不同缺氧濃度下HaCaT細胞中HSulf-1,HIF-1α蛋白的表達;構(gòu)建重組質(zhì)粒p CDH-HSulf-1;Western blot法篩選HSulf-1過表達穩(wěn)轉(zhuǎn)HaCaT細胞株。結(jié)果 HSulf-1在缺氧條件下低表達,與HIF-1α表達呈負相關(guān);成功篩選過表達HaCaT穩(wěn)定細胞株。結(jié)論 HSulf-1低表達,與腫瘤細胞或腫瘤組織中表達一致,過表達可抑制腫瘤細胞增殖。推測過表達HSulf-1可能抑制角質(zhì)形成細胞的增殖,構(gòu)建HSulf-1基因過表達載體,便于進一步體內(nèi)外研究與角質(zhì)形成細胞增殖的影響。
[Abstract]:Objective to investigate the expression of HSulf-1,HIF-1 偽 in human keratinocyte line HaCaT under hypoxia and to construct HSulf-1 gene overexpression vector. Methods Western blot assay was used to detect the expression of HSulf-1,HIF-1 偽 protein in HaCaT cells under different hypoxia concentrations, and the recombinant plasmid p-CDH-HSulf-1;Western blot was constructed to screen the stably transfected HaCaT cell lines with HSulf-1 overexpression. Results the low expression of HSulf-1 was negatively correlated with the expression of HIF-1 偽 under hypoxia, and the stable cell lines expressing HaCaT were successfully screened. Conclusion the low expression of HSulf-1 is consistent with the expression in tumor cells or tissues, and over-expression can inhibit the proliferation of tumor cells. It is speculated that overexpression of HSulf-1 may inhibit the proliferation of keratinocytes and construct HSulf-1 gene overexpression vector, which is convenient for further study in vitro and in vivo and the effect of keratinocyte proliferation.
【作者單位】: 安徽醫(yī)科大學上海皮膚病臨床學院;上海市皮膚病醫(yī)院;
【基金】:國家自然科學基金資助項目(81360236)
【分類號】:R364.4
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本文編號:2440060

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