爪哇伪枝藻胞外多糖诱导皮肤癌细胞(A431)凋亡的研究
ON APOPTOSIS OF HUMAN EPIDERMOID CARCINOMA A431 CELLS INDUCED BY THE EXTRACELLULAR POLYMERIC SUBSTANCES OF SCYTONEMA JAVANICUM
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摘要: 为探讨爪哇伪枝藻胞外多糖(Extracellular polymeric substances of Scytonema javanicum , EPS)诱导人表皮癌A431细胞凋亡及其对凋亡相关基因caspase-3、bcl-2和bax表达的影响,本实验利用MTT法检测细胞生长抑制情况;HE染色法及透射电镜进行形态学观察;单细胞凝胶电泳法(SCGE/彗星电泳)分析DNA受损情况;免疫组织化学法检测细胞内caspase-3、bcl-2和bax表达水平。结果显示EPS能显著抑制A431细胞增殖,并呈时间和剂量依赖性,作用96h的半数抑制浓度IC50为4.25mg/mL,并出现细胞凋亡的形态学改变;彗星电泳结果与对照相比6mg/mL EPS作用48h能引起A431细胞DNA严重损伤;免疫组织化学检测发现6mg/mLEPS作用72h能显著上调A431细胞内凋亡相关基因caspase-3和bax的表达,而下调bcl-2的表达。Abstract: To investigate the effect of extracellular polymeric substances of Scytonema javanicum(EPS) on apoptosis of A431 cells and the expression level of relative genes,cell viability was measured by MTT method,morphologic changes were observed by hematoxylin and eosin staining and transmission electron microscopy,DNA damage was detected by single cell gel electrophoresis(SCGE/comet assay),the expression level of caspase-3,bcl-2 and bax were detected by immunohistochemical staining.EPS significantly inhibited the proliferation of the A431 cells and the inhibitory effect was dose-and time-dependent,which was accompanied by the appearance of morphologic characteristics of apoptosis.The IC50 of A431 cells is 4.25mg/mL after the treatment of EPS for 96h.After 6 mg/mL EPS treatment for 48h,the DNA damaged seriously in A431 cells when compared with the untreated controls.The expression level of caspase-3 and bax were up-regulation while bcl-2 was down-regulation after 6mg/mL EPS treatment for 72h when compared with the untreated controls respectively.It concluded that EPS may inhibit the proliferation and induce apoptosis of A431 cells,in which caspase-3,bcl-2 and bax were involved.
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Keywords:
- Scytonema javanicum /
- Extracellular polymeric substances /
- A431 /
- Apoptos is
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