临床肿瘤学杂志

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重组人血管内皮抑素与紫杉醇联合诱导人食管癌细胞Eca-109凋亡的实验研究

孔 霞1,王秀美1,隋爱华2,刘圆圆1   

  1. 1 266003 山东青岛 青岛大学医学院附属医院肿瘤治疗中心 2 2266003 青岛大学医学院附属医院中心实验室
  • 收稿日期:2012-11-14 修回日期:2012-12-28 出版日期:2013-03-31 发布日期:2013-03-31
  • 通讯作者: 王秀美

A experimental study of recombined human endostatin in combination with paclitaxel inducing apoptosis of esophageal cancer cell line Eca-109

KONG Xia,WANG Xiumei,SUI Aihua,LIU Yuanyuan   

  1. Cancer Center,the Affiliated Hospital of College of Qingdao University,Qingdao 266003,China
  • Received:2012-11-14 Revised:2012-12-28 Online:2013-03-31 Published:2013-03-31
  • Contact: WANG Xiumei

摘要: 目的 探讨重组人血管内皮抑素(恩度)与紫杉醇联合作用对人食管癌细胞Eca-109增殖和凋亡的影响。方法 MTT法检测恩度与紫杉醇联合用药48h后对Eca-109细胞增殖的抑制作用;倒置显微镜下观察恩度、紫杉醇单药与联合作用于人食管癌Eca-109细胞48h后细胞形态学的改变;Annexin V/PI双染流式细胞术检测恩度、紫杉醇不同用药组作用48h后的细胞凋亡率;RT-PCR法检测凋亡相关基因Bcl-2、Bax、p21、p53 mRNA的表达情况。结果恩度、紫杉醇作用于Eca-109细胞48h后的半数抑制浓度(IC50)分别为276.6627μg/ml和3.8789μg/ml。恩度单药组、紫杉醇单药组、恩度+紫杉醇组、恩度→紫杉醇组、紫杉醇→恩度组作用Eca-109细胞48h后的抑制率分别为(33.62±2.30)%、(49.14±1.45)%、(56.29±0.71)%、(41.88±1.23)%和(51.48±0.98)%;与阴性对照组比较,恩度、紫杉醇各加药组对Eca-109细胞均有明显的抑制作用(P<0.01);恩度+紫杉醇组的抑制率高于其他各组(P<0.05)。光镜下恩度、紫杉醇作用48h后Eca-109细胞具有典型的凋亡形态学改变,恩度单药组、紫杉醇单药组、恩度+紫杉醇组、恩度→紫杉醇组、紫杉醇→恩度组的总凋亡率分别为(8.78±0.19)%、(13.82±0.15)%、(18.88±0.29)%、(11.37±0.24)%和(14.88±0.34)%;恩度、紫杉醇各加药组的总凋亡率均高于阴性对照组(P<0.05);恩度+紫杉醇组的总凋亡率明显高于其他各组(P<0.01)。与阴性对照组比较,恩度单药组、紫杉醇单药组、两药序贯组的Bcl-2 mRNA表达均明显降低;恩度与紫杉醇联合用药组的Bax mRNA表达均降低;各用药组的p21 mRNA表达均降低;恩度与紫杉醇序贯用药组的p53表达降低。结论 恩度与紫杉醇联合可协同诱导人食管癌细胞的凋亡,其作用机制可能与下调凋亡抑制基因的表达有关。

Abstract: Objective To investigate the combining effect of the recombined human endostatin(endostar)and paclitaxel on the proliferation and apoptosis of human esophageal carcinoma Eca-109 cells. Methods MTT was used for detecting inhibitory rates of different concentrations of endostar and paclitaxel of monotherapy and combination therapy on the Eca-109 cells at 48h;the shape changes of those cells were observed under the optical microscope;flow cytometry method was used for the detection of apoptosis rates of different groups after stained with Annexin V/PI at 48h;RT-PCR was used to detect the different expression of mRNA for apoptosis related genes(Bcl-2, Bax, p21 and p53) among different groups at 48h. Results The half inhibitory concentration (IC50) at 48h of endostar and paclitaxel on the Eca-109 cells were 276.6627μg/ml and 3.8789μg/ml. Inhibition rates of the endostar group,paclitaxel group,the concurrently medicated group,the endostar followed by paclitaxel group and the paclitaxel followed by endostar group were (33.62±2.30)%,(49.14±1.45)%,(56.29±0.71)%,(41.88±1.23)% and (51.48±0.98)%. The cell inhibition rate of endostar and paclitaxel medicating concurrently was higher than other groups(P<0.05). Under optical microscope,those medicated cells had typical apoptotic morphological changes. The apoptosis rates of endostar group, paclitaxel group, the concurrent group, endostar followed by paclitaxel group and the paclitaxel followed by endostar group were (8.78±0.19)%,(13.82±0.15)%,(18.88±0.29)%,(11.37±0.24)% and (14.88±0.34)%,respectively. The differences among the groups and compared with the negative control group were all statistically significant(P<0.05). Compared with the control group,the endostar group and two sequential groups significantly decreased mRNA expression of Bcl-2,and the paclitaxel group also reduced the mRNA expression of Bcl-2. The three ways of drug combination all presented a decreasing mRNA expression of Bax. Every group could decrease the expression of p21. Sequential medication groups reduced the expression of p53. Conclusion Endostar in combination with paclitaxel can synergisticly induce apoptosis of the human esophageal carcinoma cell,and the mechanism may be related to the reduced expressions of antiapoptosis genes.

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