摘要目的 通过研究PAR-1对肺巨细胞癌细胞高、低转移细胞株PLA801D和PLA801C[Ca~(2+)]I的影响,探讨PAR-1参与调节肺癌细胞转移相关功能的分子机制.方法 激光共聚焦显微镜检测PLA801D和PLA801C细胞[Ca~(2+)]I;将反义和正义PAR-1分别转染至PLA801D(D-)和PLA801C(C+)中;用thrombin和TRAP激活PAR-1,观察PAR-1对D-和C+的[Ca~(2+)]I影响.结果 PLA801D(荧光强度59.55,下同)和PLA801C(35.46)细胞之间的平均[Ca~(2+)]I差异有统计学意义(P<0.01).C+的[Ca~(2+)]I(45.77)明显高于其对照组CV(35.46,P<0.05),D-的[Ca~(2+)]I(48.42)明显低于对照组DV(59.55,P<0.05).C+和CV的[Ca~(2+)]I分别在加入thrombin 80 s和100 s后荧光强度分别达峰值48.19±9.84和45.64±9.87(P<0.05);二者均在加入TRAP60 s后达峰值,分别为111.31±25.00和52.93±11.21(P<0.05).D-和DV的[Ca~(2+)]I在加入thrombin 60 s后达峰值,分别为40.71±5.89和61.07±21.36(P<0.05),二者均在加入TRAP 40 s后达峰值,分别为84.98±11.23和102.58±21.48(P<0.05).结论 PLA801D和PLA801C的转移潜能可能与其细胞的[Ca~(2+)]I有关.激活PAR-1能够上调PLA801D和PLA801C的细胞内Ca~(2+)信号.PAR-1促进肺巨细胞癌转移的机制与上调细胞内Ca~(2+)有关.

abstractsObjectives To investigate molecular mechanisms of PAR-1 regulation on intracellular Ca~(2+) mobilization in lung giant cell carcinoma cells in vitro and its involvement in tumor metastasis.Methods Free intracellular Ca~(2+)([Ca~(2+)]i) was measured in lung giant cell carcinoma PLA801C and PLA801D cells by confocal microscopy. Sense and anti-sense PAR-1 expression vectors were transfected into PLA801C (C+) and PLA801D(D-) cells, respectively. The effects of PAR-1 expression were investigated by thrombin and TRAP-induced mobilization of [Ca~(2+)]i in the C+ and D-cells. Results There were significant differences of the mean values of [Ca~(2+)]i between PLA801D (59.55) and PLA801C cells (35.46, P<0.01). The mean [Ca~(2+)]i of C+ cells (45.77) was significantly higher than that of its control CV cells (35.46, P < 0.05), and the mean [Ca~(2+)]i of D-cells (48.42)was significantly lower than that of its control DV cells (59.55, P<0.05). The peaks of [Ca~(2+)]i of C+ and CV cells were 48. 19±9.84 and 45.64±9.87 (P<0.05)respectively at 80 s and 100 s after thrombin treatment, but were 111.31±25.00 and 52.93±11.21 (P < 0. 05) respectively at 60 s after TRAP treatment. The peaks of[Ca~(2+)]i of D-and DV cells were 40. 71±5.89 and 61.07±21.36 (P < 0.05) respectively at 60 s after thrombin treatment,but were 84. 98±11.23 and 102.58±21.48 (P<0.05) respectively at 40 s after TRAP treatment. Conclusions The high metastatic potential of PLA801D and PLA801C may be related to [Ca~(2+)]i of the tumor cells. PAR-1 may play an important role in the metastasis of lung giant cell carcinoma cells by up-regulating the intracellular Ca~(2+).