[Exogenous expression of SOCS box-deficient mutant ASB-8 suppresses the growth of lung adenocarcinoma SPC-A1 cells].

Yong-Zhong Liu, Jin-Jun Li, Feng-Rui Zhang, Yan Shen, Ming Yao, Da-Fang Wan, Jian-Ren Gu
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Abstract

ASB-8 is a new member of the human ankyrin repeat and SOCS box containing protein family (ASB). This report deals with the expression of ASB-8 protein in lung carcinoma tissue, as well as its biological effect on the proliferation and growth of lung cancer cell line SPC-A1. ASB-8 was expressed in pT7-450 expression vector, and an anti-ASB-8 rabbit polyclonal antibody was prepared. The expression of ASB-8 protein in lung carcinoma tissue was detected using immunohistochemistry. The growth characteristics of the lung adenocarcinoma SPC-A1 cells expressing either exogenous ASB-8 or ASB-8 SB (SOCS box-deficient) was studied by both in vitro cell growth curve and in vivo nude mouse tumor formation assay. Immunohistochemical staining detected positive reaction of 96.8% (30/31) showing that ASB-8 was highly expressed in lung cancer tissues; however, the expression of ASB-8 was lower, or even no expression in noncancerous lung tissues. Significant growth inhibition was observed in SPC-A1 cell line expressing ASB-8 SB on day 4 and persisted to day 6, compared with mock transfected cells (P<0.01). No difference in the growth properties was observed between ASB-8 and mock transfected cells. In vivo study also showed that tumor formation of ASB-8 SB expressing cells was significantly inhibited as compared with that of the control group (P<0.01). Therefore, ASB-8 may play an important role in growth and proliferation of lung cancer, possibly as positive regulator.

[外源表达SOCS盒缺陷突变体ASB-8抑制肺腺癌SPC-A1细胞的生长]。
ASB-8是人类锚蛋白重复序列和含SOCS盒蛋白家族(ASB)的新成员。本文研究ASB-8蛋白在肺癌组织中的表达及其对肺癌细胞系SPC-A1增殖生长的生物学作用。将ASB-8在pT7-450表达载体上表达,制备抗ASB-8兔多克隆抗体。采用免疫组化方法检测肺癌组织中ASB-8蛋白的表达。采用体外细胞生长曲线和体内裸鼠成瘤实验研究了外源性ASB-8或ASB-8 SB (SOCS box-deficient)表达的肺腺癌SPC-A1细胞的生长特性。免疫组化染色检测到96.8%(30/31)的阳性反应,表明ASB-8在肺癌组织中高表达;而ASB-8在非癌性肺组织中表达较低,甚至不表达。与模拟转染的细胞相比,表达asb - 8sb的SPC-A1细胞系在第4天观察到明显的生长抑制,并持续到第6天
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