基質交互分子1是鈣池調控鈣離子流重要的一員，藉以調控細胞內鈣離子濃度以及相關之功能。近來研究指出血小板生長因子可在細胞內產生內質網鈣離子流出並透過基質交互分子與鈣釋放激活通道所形成之鈣離子通道造成鈣池調控鈣離子流。因此，本篇研究有興趣於基質交互分子於血小板生長因子BB型引發之鈣離子震盪波及其相關功能。過去研究指出血小板生長因子引發鈣離子上游主要透過PLCγ途徑，活化的PIP2可被水解為DAG和IP3並造成內質網鈣離子流出，進而內質網鈣離子缺失引發鈣池調控鈣離子流活化。此外，血小板生長因子亦可活化其他訊息途徑，如AKT、JNK、ERK和STAT3。本研究藉由多功能微量盤分光光譜儀分析，我們發現使用高濃度的血小板生長因子(100 ng/mL)可以引發明顯之鈣離子上升，而此上升主要存在於基質交互分子1剃除之小鼠胚胎上皮細胞中而非野生種。免疫墨點分析結果發現在於基質交互分子剃除之小鼠胚胎上皮細胞中，血小板生長因子受體有增強表達的現象，並造成下游PLCγ、AKT、JNK、ERK等途徑增強之活化表現。本研究亦藉由鈣池調控鈣離子流、PLC、血小板生長因子受體之抑制劑，可明顯抑制血小板生長因子所引發之鈣離子上升和蛋白質磷酸化表現。免疫螢光分析基質交互分子2在缺乏基質交互分子1時取代其原有的功能，透過血小板生長因子導致內質網鈣離子缺失時活化，引發規模較小之鈣質調控鈣離子流。綜合以上實驗數據可以得知，在基質交互分子剔除之小鼠胚胎上皮細胞中，血小板生長因子受體蛋白有增強的表現，其下游引發增強之PDGFR-PLCγ-STIM2途徑，使得相對於野生種有明顯的鈣離子上升。

STIM1 (stromal interaction molecule 1) has been reported as a key molecule in store-operated calcium entry (SOCE) which is one of the major mechanisms that allows extracellular Ca2+ refill into cytoplasm to regulate many cell functions. Platelet-derived growth factor-BB (PDGF-BB) induces cell migration and proliferation in MEF (murine embryo fibroblast) cells. Recent studies have suggested that PDGF-BB induces endo-plasm reticulum (ER) Ca2+ release and SOCE through the STIM1-Orai1 complex. In this study, we investigated the role of STIM1 in PDGF-BB-induced Ca2+ oscillation and its functions in MEF cells. PDGF-BB activates Ca2+ elevation by activating the PLCγ sig-naling pathway, and the PIP2 hydrolysis into IP3 and DAG resulted in ER Ca2+ release. The depletion of ER Ca2+ induces SOCE through forming a SOC channel (STIM-TRPC-Orai complex). Also, there are several signaling pathways co-activated by PDGF-BB stimulation, including AKT, JNK, ERK and STAT3. According to data collect-ed from the FlexStation3 microplate reader, unexpectedly, we found PDGF-BB (100 ng/ml) induced manifest Ca2+ elevation in STIM1 knockout MEF cells dramatically with different doses and immunoblots data also indicated that PDGFR (PDGFα and PDGFRβ) overexpression induced PLCγ activity time-dependently with PDGF-BB stimulation in STIM1 knockout MEF cells. Furthermore, we applied SOCE, PLC, PDGFR inhibitors to examine the signaling transduction pathway. 2-APB, SKF96365, YM58483, Gd3+, La3+ were applied to inhibit PDGF-BB-induced Ca2+ elevation, U73122 and D609 were applied to inhibit PLC phosphorylation, and the AG-1295 was applied to inhibit PDGFR activa-tion after PDGF-BB triggering. These data show that PDGFR protein levels are enhanced in STIM1 knockout MEF cells and then raise the PDGF-PLCγ-STIM2 signaling cascade to increase the [Ca2+]i with PDGF-BB stimulation.

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