nm23-H1基因转染对人高转移大细胞肺癌细胞株生物学行为的影响及作用机理

The mechanism and influence on the biological behavior of human high-metastatic large cell lung cancer cell lines with transfection of nm23-H1 gene

背景与目的nm23-H1基因已被证明是一个肿瘤转移抑制基因,但其相关作用机理仍不明确。本研究通过比较nm23-H1基因转染前后人高转移大细胞肺癌细胞株L9981生物学行为的变化,探讨nm23-H1基因影响人高转移大细胞肺癌细胞株生物学行为的可能机理。方法应用细胞体外增殖活性检测(MTT法)和体外侵袭力检测(改良Boyden小室法)技术分别检测nm23-H1基因转染前后人高转移大细胞肺癌细胞株L9981、L9981-pLXSN和L9981-nm23-H1肺癌细胞株体外增殖活性和侵袭力的变化;同时加入PKC特异抑制剂Calphostin C作用后,再次观察三株细胞株抑制剂作用前后细胞侵袭力、增殖力的变化。结果nm23-H1基因缺失的人高转移大细胞肺癌细胞株L9981和L9981-pLXSN体外侵袭力和增殖活性明显高于转染nm23-H1基因的L9981-nm23-H1肺癌细胞株(P<0.001);L9981和L9981-pLXSN细胞间体外侵袭力和增殖活性则无统计学差异(P>0.05)。用PKC抑制剂Calphostin C处理L9981、L9981-pLXSN和L9981-nm23-H1肺癌细胞株后,细胞体外侵袭力和增殖活性下降(P<0.001),而转染nm23-H1基因的L9981-nm23-H1细胞体外侵袭力和增殖活性仍低于L9981和L9981-pLXSN(P<0.001),L9981和L9981-pLXSN细胞间则无统计学差异(P>0.05)。结论nm23-H1基因可明显抑制L9981肺癌细胞株的细胞增殖力和侵袭力。nm23-H1基因对人高转移大细胞肺癌细胞株L9981的生物学行为的影响可能与其抑制PKC信号传导有关。

Background and objective It has been confirmed that nm23-H1 gene is one of the tumor metastasis suppressor genes. Up to now, the exact mechanism of nm23-H1 gene is uncertian. The aim of this study is to compare the biological behavior changes among three human high-metastatic large cell lung cancer cell lines which transfected and untransfeeted nm23-H1 gene, and to study the mechanism of nm23-H1 gene supressing the metastasis. Methods Boyden Chamber and MTT method were used to detect the rates of invasion and proliferation among different human pulmonary carcinoma cells of transfected and untransfected nm23-H1 gene; meanwhile The three lung cancer cell lines were treated with PKC inhibitor Calphostin C, and the above measurements were also performed. Results The ability of invasion and proliferation of L9981 and L9981-PLXSN human high-metastatic large cell lung cancer cells,which lack of nm23-H1gene, was higher than that of L9981-nm23-H1 human high-metastatic large cell line, which transfected with nm23-Hlgene （P〈0.001）. There was no difference heteween L9981 and L9981-PLXSN cell lines （P〉0.05）. After treated with PKC inhibitor Calphstin C, the invasion and proliferation ability of three lung cancer cell lines were obviously go down （P〈0.001）, however, the invasion and proliferation ability of L9981-nm23-H1 lung cancer cell line was still lower than those of L9981 and L9981-PLXSN lung cancer cell lines （P〈0.001）, and there was also no significant difference between two later cell lines （P〉0.05）. Conclusion Our data suggest that nm23-H1 gene can significantly inhibit the cell proliferation and invasion in L9981 lung cancer line. The effect of nm23-H1 might be correlated with downregulation of PKC signal transduction in human high-metastatic large cell lung cancer cell line.