Nie Qiang, Zhu Wen, Liu Lunxu, Fu Junke, Li Dingbiao, Li Yin, Chen Jun, Wu Zhihao, Zhou Qinghua
Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenviroment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Anshan Road No.154, Heping District, Tianjin 300052, China;Guangdong Provincial People's Hospital, Guangzhou, Guangzhou 510080, China.
Zhongguo Fei Ai Za Zhi. 2008 Jun 20;11(3):363-7. doi: 10.3779/j.issn.1009-3419.2008.03.034.
Protein Kinase C (PKC) is one of the key kinases in the cell signal transduction passway. There are more reports about it's ability on cell proliferation, but fewer on invasion and metastasis in the past; and it's mechanisms are unclear. The aim of this study is to analyze the variation of intracellular distribution and translocation of the protein kinase C alpha among human high-metastatic large cell lung cancer cell line with different metastasis potential,in order to investigate the correlation between the lung carcinoma invasion and metastasis and the PKC isoforms.
Using Western blot and laser scanning confocal microscope (LSCM) method. The distribution of PKC alpha in cytosol and plasma membrane and translocation were detected among different metastatic potential human pulmonary carcinoma cells L9981, L9981-pLXSN and L9981-nm23-H1 before and after treatment with PKC inhibitor Calphostin C, by Western blot and LSCM.
PKC alpha in L9981 and L9981- pLXSN was mainly expressed on membrane, which was remarkably higher than those in L9981-nm23-H1 cell line (P =0.001); while expression of PKC alpha in cytoslol in L9981 and L9981-pLXSN cell lines, was lower than those in L9981-nm23-H1 cell line (P <0.001). The expression of PKC alpha in cytosol in L9981-nm23-H1 cell line was remarkably higher than those in L9981 and L9981-pLXSN cell lines (P <0.001), while expression of PKC alpha in plasma membrane in L9981-nm23-H1 cell line, was significantly lower than those in L9981 and L9981- pLXSN cell lines (P =0.001). PKC alpha is mainly located in nucleus and perinucleus in L9981 and L9981-pLXSN cells, which was in active status, In L9981-nm23-H1 cell line, PKC alpha is mainly located in soluble cytosolic section, which was in inactive status. After treated with PKC inhibitor Calphostin C, the expression of PKC alpha in membrane in L9981, L9981-pLXSN and L9981-nm23-H1 was downregulated, and PKC alpha were observed mainly located in cytosolic site in all the three cell lines, which was mainly in inactive status.
The study suggests that PKC isoforms is closely correlated with human lung cancer invasion and metastasis.
蛋白激酶C（PKC）是细胞信号转导通路中的关键激酶之一。过去关于其细胞增殖能力的报道较多，而关于侵袭和转移的报道较少，其机制尚不清楚。本研究旨在分析不同转移潜能的人高转移大细胞肺癌细胞系中蛋白激酶Cα的细胞内分布和转位变化，以探讨肺癌侵袭转移与PKC亚型之间的相关性。
采用蛋白质印迹法和激光扫描共聚焦显微镜（LSCM）方法。用PKC抑制剂Calphostin C处理前后，通过蛋白质印迹法和LSCM检测不同转移潜能的人肺癌细胞L9981、L9981-pLXSN和L9981-nm23-H1中PKCα在细胞质和质膜中的分布及转位情况。
L9981和L9981-pLXSN中的PKCα主要表达于细胞膜上，显著高于L9981-nm23-H1细胞系（P =0.001）；而L9981和L9981-pLXSN细胞系细胞质中PKCα的表达低于L9981-nm23-H1细胞系（P <0.001）。L9981-nm23-H1细胞系细胞质中PKCα的表达显著高于L9981和L9981-pLXSN细胞系（P <0.001），而L9981-nm23-H1细胞系质膜中PKCα的表达显著低于L9981和L9981-pLXSN细胞系（P =0.001）。PKCα在L9981和L9981-pLXSN细胞中主要位于细胞核和核周，处于激活状态；在L9981-nm23-H1细胞系中，PKCα主要位于可溶性细胞质部分，处于失活状态。用PKC抑制剂Calphostin C处理后，L9981、L9981-pLXSN和L9981-nm23-H1细胞膜中PKCα的表达下调，在这三种细胞系中均观察到PKCα主要位于细胞质部位，主要处于失活状态。
该研究提示PKC亚型与人肺癌侵袭转移密切相关。
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