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LAPTM4B 的过表达促进胆囊癌细胞在体外的生长。

Overexpression of LAPTM4B promotes growth of gallbladder carcinoma cells in vitro.

机构信息

Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing, China.

出版信息

Am J Surg. 2010 Apr;199(4):515-21. doi: 10.1016/j.amjsurg.2009.03.031. Epub 2009 Dec 2.

Abstract

BACKGROUND

The overexpression of LAPTM4B-35 in gallbladder carcinoma (GBC) and its clinicopathologic and prognostic significance have been previously shown. Thus, this gene may play a role in the growth of GBC cells.

METHODS

The pcDNA3-AE containing the complete open reading frame of LAPTM4B (lysosome-associated protein transmembrane-4beta) and mock (pcDNA3) plasmids were transiently transfected into GBC-SD cells. Cell proliferation, cell cycle distribution, and protein expression were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium assay, flow cytometry, and Western blot, respectively.

RESULTS

Cells transfected with pcDNA3-AE revealed accelerated proliferation, less serum dependence, and significant cell cycle progression compared with cells transfected with mock plasmid and parent cells. These phenotypes were accompanied by upregulated expression of C-myc, c-Fos, c-Jun, cyclin D1, and cyclin E and downregulated expression of P16 and P-27.

CONCLUSIONS

LAPTM4B overexpression promotes the growth of GBC cells in vitro by regulating the expression levels of some proliferation-associated proteins. Therefore, the LAPTM4B gene might be used as a novel therapeutic target of GBC.

摘要

背景

先前已经表明,LAPTM4B-35 在胆囊癌(GBC)中的过表达及其与临床病理和预后的关系。因此,该基因可能在 GBC 细胞的生长中发挥作用。

方法

瞬时转染含有完整 LAPTM4B(溶酶体相关蛋白跨膜-4β)开放阅读框的 pcDNA3-AE 和空载(pcDNA3)质粒至 GBC-SD 细胞。通过 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑蓝比色法、流式细胞术和 Western blot 分别评估细胞增殖、细胞周期分布和蛋白质表达。

结果

与转染空载质粒和亲本细胞的细胞相比,转染 pcDNA3-AE 的细胞显示出增殖加速、对血清依赖性降低和明显的细胞周期进展。这些表型伴随着 C-myc、c-Fos、c-Jun、细胞周期蛋白 D1 和细胞周期蛋白 E 的表达上调,以及 P16 和 P-27 的表达下调。

结论

LAPTM4B 的过表达通过调节一些与增殖相关的蛋白质的表达水平,促进 GBC 细胞在体外的生长。因此,LAPTM4B 基因可能被用作 GBC 的新的治疗靶点。

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