Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing, China.
Am J Surg. 2010 Apr;199(4):515-21. doi: 10.1016/j.amjsurg.2009.03.031. Epub 2009 Dec 2.
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.
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.
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.
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.
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。
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。
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。
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。
