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研究地塞米松(DEX)对顺铂(CDDP)引起的人肺腺癌细胞SPC-A1凋亡的抑制作用及可能的分子机制。将不同浓度的DEX分别加入人肺腺癌细胞SPC-A1体外诱导培养24h后,再用不同浓度CDDP处理SPC-A1细胞48h。MTT法检测细胞存活率;RT-PCR方法检测1μmol/L DEX诱导培养不同时间后SPC-A1细胞内血清/糖皮质激素-诱导激酶(SGK-1)和有丝分裂原蛋白激酶(MKP-1)的表达;用生物素标记的抗糖皮质激素受体(GR)抗体对SPC-A1细胞行免疫组化染色来检测GR的表达。MTT测定结果显示,SPC-A1细胞经DEX诱导后,对CDDP所致的凋亡有抵抗作用并与DEX浓度呈剂量依赖关系。RT-PCR检测到DEX诱导培养能提高SGK-1在SPC-A1细胞中的表达,表达量随时间延长而增加;但未检测到MKP-1的表达。免疫组化检测显示SPC-A1细胞经DEX诱导后GR上调,细胞内GR表达的阳性细胞数明显高于对照组。结果表明DEX对CDDP引起SPC-A1细胞的凋亡有抑制作用。可能的分子机制是通过上调细胞内GR表达,进而上调其通路下游抗凋亡蛋白SGK-1的表达,导致SPC-A1细胞产生抗凋亡作用。
To investigate the inhibitory effect of dexamethasone (DEX) on the apoptosis of human lung adenocarcinoma cell line SPC-A1 induced by cisplatin (CDDP) and its possible molecular mechanisms. After different concentrations of DEX were added to human lung adenocarcinoma cell line SPC-A1 for 24 hours respectively, SPC-A1 cells were treated with different concentrations of CDDP for 48 hours. MTT assay was used to detect the cell viability. The expression of SGK-1 and MKP-1 in SPC-A1 cells induced by 1 μmol / L DEX was detected by RT-PCR. The expression of GR was detected by immunohistochemistry staining of SPC-A1 cells with anti-glucocorticoid receptor (GR) antibody labeled with biotin. MTT assay showed that after induced by DEX, SPC-A1 cells were resistant to CDDP-induced apoptosis and in a dose-dependent manner with DEX concentration. The expression of SGK-1 in SPC-A1 cells was detected by RT-PCR after DEX induction. The expression of SGK-1 in SPC-A1 cells increased with time. However, the expression of MKP-1 was not detected. Immunohistochemistry showed that GR was upregulated in SPC-A1 cells induced by DEX, and the number of GR positive cells in the cells was significantly higher than that in the control group. The results show that DEX can inhibit the apoptosis of SPC-A1 cells induced by CDDP. The possible molecular mechanism is through the upregulation of intracellular GR expression, which in turn up-regulates the expression of its downstream anti-apoptotic protein SGK-1, resulting in anti-apoptotic effects of SPC-A1 cells.