壳寡糖对Hela细胞的增殖抑制与诱导凋亡

Effects of chitooligosaccharide(COS) on proliferation inhibition and apoptosis in Hela cell line

为了明确壳寡糖(COS)对Hela细胞的增殖抑制作用及对凋亡的影响,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测增殖抑制作用,Wrights-Giemsa及吖啶橙-溴乙锭(AO-EB)染色观察凋亡细胞的形态,Annexin V-FITC/PI法检测凋亡率,免疫组化法分析Bax,Bcl-2和Survivin表达量变化.结果显示:不同浓度COS处理Hela细胞均能抑制增殖,5.0mg/mL并作用24h对其增殖抑制率为52.15%(P<0.01).Wrights-Giemsa染色显示细胞形态趋近圆形,贴壁能力减弱,出现凋亡小体.AO-EB染色时细胞出现早期和晚期凋亡现象.AnnexinV-FITC/PI检测细胞凋亡率为31.75%(P<0.05).免疫组化分析细胞内Bax表达量增加,Bcl-2和Survivin表达量降低.表明COS能够抑制Hela细胞增殖并诱导凋亡,其原因与Bax的上调和Bcl-2与Survivin的下调相关.

To explore the effect of inhibiting proliferation and the mechanism of apoptosis induced by chitooligosaccharide（COS） on Hela, 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide （MTT） assay was used for evaluation of proferation, the morphological changes of cells using Wright＇s- Giemsa and AO-EB staining were observed, Annexin V-FITC/PI double staining was employed for quanti- fying apoptotic cells by flow cytometry and the expression level of Bax, Bcl-2, Survivin in Hela were detec- ted by immunohistochemistry, respectively. The results showed that different concentrations of COS could inhibite proliferation of Hela. After treated with 5.0 mg/mL COS for 24 h, there was a maximum （up to 52. 15 % , P〈0.01）. Wright＇s-Giemsa staining showed that cell morphology became round and some cells floated above the wall also apoptosis cells in early phase and late phase were observed by AOEB staining. Annexin V FITC/PI quantitative detection the apoptosis rate of cells was 31.75 % （P〈0.05）. The expres- sion level of proapoptotic protein Bax was increased by immunohistochemistry, however, the expressionlevel of antiapoptotic protein Bcl-2 and Survivin was decreased. In conclusion,COS can induce proliferation inhibition and apoptosis. The mechanism of apoptosis induced by COS was relative to the expression level of Bax, Bcl-2, and Survivin.