MAG-1对人肺癌细胞转移能力的影响及其在24例肺癌组织中的表达

Promotion of MAG-1 on Metastasis of Lung Cancer Cells in vitro and Its Expression in Lung Cancer Tissue of 24 Cases

背景与目的肺癌转移相关基因-1(Pulmonary cancer Metastasis Associated Gene-1,MAG-1)是利用抑制消减杂交技术获得的与肺癌转移相关的基因。此研究的目的在于探讨基因MAG-1对肺巨细胞癌细胞的转移相关生物学行为的影响及其在24例肺癌组织中的表达情况。方法构建MAG-1的正反义表达载体,分别转染PLA801C和PLA801D,获得稳定表达相应序列的细胞株,观察稳定转染细胞的多种肿瘤转移相关能力的改变,并检测MAG-1对CD44、MMP-2等分子的影响,最后还检测了MAG-1在源于不同转移状况患者的肺癌标本中的表达情况。结果MAG-1对细胞生长、细胞与胞外基质的粘附能力以及侵袭能力均有促进作用;并能促进细胞中CD44和MMP-2的表达;MAG-1mRNA在有转移的肺癌中的阳性率(7/12)高于其在无转移的肺癌标本中的阳性率(2/12)。结论MAG-1具有增强肺癌细胞转移能力的功能,并且在转移性肺癌组织中其mRNA具有较高的阳性检出率,是可能的肺癌转移相关基因。

Background and objective Tumor metastasis is a muhistep process with many genes involved in. A novel gene MAG-1, identified by suppression subtractive hybridization from lung cancer cells was found to be associated with tumor metastasis. The aims of this work are to investigate the metastasis related effects of MAG-1 on human lunggiant-cell line PLA801, and to compare the expression rate of MAG-1 in cancer tissue from lung cancer patients with different metastatic status. Methods Sense and anti-sense expressing vectors of MAG-1 were constructed and transfected into PLA801C and PLA801D respectively. Colony forming, adherence assay, MTT assay and Transwell experiments were used to evaluate the alterations of clone forming, cell-matrix adherence, proliferating and invasion of the stable transfected cell strains. Western blot was employed to detect the proteins levels of CD44, and MMP-2 in cell strains and mRNA state of MAG-1 in lung cancer tissue from patients with or without pathological metastasis were also analyzed by RTPCR. Results MAG-1 could increase cell-ECM （Extracellular Matrix） adhesive capacity, promote invasion, enhance cell proliferation and had no effects on clone forming ability of PLA801 cells in soft agar. MAG-1 was also found have positive effects on the protein level of CD44 and MMP-2 in PLA801 cells, and the detection rate of MAG-1 mRNA was much higher in cancer tissue from metastatic patients （7/12） than that in non-metastatic patients （2/12）. Conclusion MAG-1 could promote lung cancer metastasis and might be a metastasis associated gene of lung cancer.