肾癌细胞与内皮细胞的黏附依赖于 PKCmu。
Adhesion of renal carcinoma cells to endothelial cells depends on PKCmu.
机构信息
Department of Urology, University Medical Center Mainz, Mainz, Germany.
出版信息
BMC Cancer. 2010 May 6;10:183. doi: 10.1186/1471-2407-10-183.
BACKGROUND
The formation of metastases includes the separation of tumor cells from the primary tumor, cell migration into subendothelial tissue and cell proliferation in secondary organ. In this process, cell adhesion of tumor cells to the endothelium is an essential requirement for formation of metastases. Protein kinase C (PKC) regulates adhesion and proliferation. To identify a relation between PKC isoforms and tumor progression in renal cell carcinoma (RCC), the influence of PKC isoforms on cell adhesion and proliferation, and possible influences of integrins were analyzed in RCC cells.
METHODS
The experiments were performed in the RCC cell lines CCF-RC1 and CCF-RC2 after pre-incubation (16 h) with the PKC inhibitors GF109203X (inhibits PKCalpha, betaI, betaII, gamma, delta and epsilon), GO6976 (inhibits PKCalpha, betaI and mu), RO31-8220 (inhibits PKCalpha, betaI, betaII, gamma and epsilon) and rottlerin (inhibits PKCdelta). Cell adhesion was assessed through adherence of RCC cells to an endothelial monolayer. Cell proliferation was analyzed by a BrdU incorporation assay. The expression of beta1 integrins was analyzed by flow cytometry.
RESULTS
In CCF-RC1 cells, cell adhesion was significantly reduced by GO6976 to 55% and by RO31-8220 to 45% of control. In CCF-RC2 cells, only GO6976 induced a significant reduction of cell adhesion to 50% of control levels. Proliferation of both cell lines was reduced by rottlerin to 39% and 45% of control, respectively. The beta1 integrin expression on the cell surface of CCF-RC1 and CCR-RC2 cells was decreased by RO31-8220 to 8% and 7% of control, respectively. beta2 and beta3 integrins were undetectable in both cell lines.
CONCLUSIONS
The combination of the PKC inhibitors leads to the assumption that PKCmu influences cell adhesion in CCF-RC1 and CCF-RC2 cells, whereas in CCF-RC1 cells PKCepsilon also seems to be involved in this process. The expression of beta1 integrins appears to be regulated in particular by PKCepsilon. Cell proliferation was inhibited by rottlerin, so that PKCdelta might be involved in cell proliferation in these cells.
背景
转移的形成包括肿瘤细胞从原发性肿瘤分离、细胞迁移到内皮下组织和在次级器官中的增殖。在这个过程中，肿瘤细胞与内皮的细胞黏附是形成转移的必要条件。蛋白激酶 C（PKC）调节黏附和增殖。为了确定蛋白激酶 C 同工型与肾细胞癌（RCC）肿瘤进展之间的关系，分析了 RCC 细胞中 PKC 同工型对细胞黏附与增殖的影响以及整合素的可能影响。
方法
在 CCF-RC1 和 CCF-RC2 的 RCC 细胞系中进行实验，这些细胞系在预孵育（16 小时）后用蛋白激酶 C 抑制剂 GF109203X（抑制 PKCalpha、betaI、betaII、gamma、delta 和 epsilon）、GO6976（抑制 PKCalpha、betaI 和 mu）、RO31-8220（抑制 PKCalpha、betaI、betaII、gamma 和 epsilon）和罗特林（抑制 PKCdelta）处理。通过 RCC 细胞与内皮单层的黏附来评估细胞黏附。通过 BrdU 掺入测定法分析细胞增殖。通过流式细胞术分析 beta1 整合素的表达。
结果
在 CCF-RC1 细胞中，细胞黏附分别被 GO6976 抑制至对照组的 55%和 RO31-8220 抑制至 45%。在 CCF-RC2 细胞中，只有 GO6976 诱导细胞黏附显著减少至对照组的 50%。两种细胞系的增殖均被罗特林抑制至对照组的 39%和 45%。CCF-RC1 和 CCR-RC2 细胞表面的 beta1 整合素表达分别被 RO31-8220 抑制至对照组的 8%和 7%。两种细胞系均未检测到 beta2 和 beta3 整合素。
结论
PKC 抑制剂的联合应用提示 PKCmu 影响 CCF-RC1 和 CCF-RC2 细胞的细胞黏附，而在 CCF-RC1 细胞中 PKCepsilon 似乎也参与了这一过程。beta1 整合素的表达似乎特别受 PKCepsilon 的调节。罗特林抑制细胞增殖，因此 PKCdelta 可能参与这些细胞的细胞增殖。
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