YKL-40调控子宫内膜癌顺铂化疗耐药性的实验研究

摘要/Abstract

摘要： 目的探讨沉默甲壳质酶蛋白 40（YKL-40）表达对子宫内膜癌顺铂（DDP）耐药细胞株Ishikawa／DDP的影响。方法采用DDP长期浓度梯度递增法体外建立Ishikawa／DDP耐药细胞株，并检测多药耐药相关基因（MDR1）和Bcl-2、Bax和caspase-3的表达，采用MTT法检测Ishikawa细胞和Ishikawa／DDP细胞的增殖活性，计算半数抑制浓度（IC50）。Ishikawa／DDP分别转染NC阴性序列（NC组）和siRNA YKL-40（si-YKL-40组），另设未转染细胞作为对照组，采用MTT法、划痕实验和Annexin V／PE双染法检测DDP对si-YKL-40组Ishikawa／DDP细胞增殖、迁移能力和凋亡的影响。结果体外成功建立Ishikawa／DDP耐药细胞株，3.125、6.25、12.5、25、50、100 μmol／L DDP对Ishikawa／DDP细胞的增殖抑制率分别为（6.93±2.45）％、（8.14±4.50）％、（11.37±4.62）％、（15.18±3.97）％、（26.29±5.08）％、（41.32±7.64）％，明显低于Ishikawa细胞（P＜0.05）。DDP对Ishikawa和Ishikawa／DDP的IC50分别为14.58 μmol／L和116.70 μmol／L，Ishikawa／DDP的耐药指数为8.004。QPCR检测结果显示，Ishikawa细胞YKL-40、MDR1、Bcl-2、Bax、caspase-3 mRNA表达量分别为0.82±0.15、0.43±0.11、1.05±0.23、1.17±0.20、0.96±0.18，而Ishikawa／DDP细胞分别为1.87±0.40、2.34±0.46、1.52±0.28、0.72±0.21、0.49±0.17，差异有统计学意义（P＜0.05）。3.125、6.25、12.5、25、50、100 μmol／L DDP对si-YKL-40组细胞的增殖抑制率分别为（10.95±2.74）％、（18.73±5.30）％、（32.79±5.47）％、（52.28±6.58）％、（61.73±5.26）％、（65.45±7.33）％，明显高于对照组和NC组，差异有统计学意义（P＜0.05）。DDP对si-YKL-40组细胞的IC50为22.19 μmol／L。与对照组和NC组比较， si-YKL-40组细胞凋亡率升高、愈合率下降；YKL-40、MDR1、Bcl-2蛋白表达量下调， Bax和caspase-3蛋白表达量上调，差异有统计学意义（P＜0.05）。结论沉默YKL-40可显著提高Ishikawa／DDP细胞株对DDP的敏感性，并促进细胞凋亡，其具体机制可能与下调MDR1、Bcl-2表达，及上调Bax和caspase-3表达有关。

关键词:

子宫内膜癌')">"> 子宫内膜癌, YKL-40, 顺铂, 耐药性

Abstract:

Objective To investigate the effect of silenced chitinase 40 （YKL-40） expression on cisplatin-resistant endometrial cancer cell line Ishikawa／DDP. Methods Ishikawa／DDP drug-resistant cell lines were established by DDP long-term concentration gradient method in vitro， and the expression of MDR1， Bcl-2， Bax and caspase-3 were detected. The proliferation activity of Ishikawa cells and Ishikawa／DDP cells was detected by MTT method， and the half inhibitory concentration （IC50） was calculated. Ishikawa／DDP was transfected with NC negative sequence （NC group） and siRNA YKL-40 （si-YKL-40 group）， respectively. The untransfected cells were set as blank control group. The effects of DDP on proliferation， migration and apoptosis of Ishikawa／DDP cells in si-YKL-40 group were detected by MTT， scratch test and Annexin V／PE double staining. Results Ishikawa／DDP resistant cell lines were successfully established in vitro. The inhibitory rates of 3.125， 6.25， 12.5， 25， 50 and 100 μmol／L DDP on proliferation of Ishikawa／DDP cells were （6.93±2.45）％，（8.14±4.50）％，（11.37±4.62）％，（15.18±3.97）％，（26.29±5.08）％，（41.32±7.64）％， which were significantly lower than those of Ishikawa cells （P＜0.05）. The IC50 were 14.58 μmol／L and 116.70 μmol／L， respectively. The resistance index of Ishikawa／DDP was 8.004. The expression of YKL-40， MDR1 and Bcl-2 in Ishikawa／DDP cells were 1.87±0.40， 2.34±0.46 and 1.52±0.28， which were higher than those in Ishikawa cells （P＜0.05）. The expression of Bax and caspase-3 in Ishikawa／DDP cells were 0.72±0.21 and 0.49±0.17， which were lower than those in Ishikawa cells （P＜0.05）. The inhibitory rates of 3.125， 6.25， 12.5， 25， 50 and 100 μmol／L DDP on the proliferation of si-YKL-40 cells were （10.95±2.74）％，（18.73±5.30）％，（32.79±5.47）％，（52.28±6.58）％，（61.73±5.26）％，（65.45±7.33）％， respectively， which were significantly higher than those of blank control group and NC group （P＜0.05）. The IC50of DDP on si-YKL-40 cells was 22.19 μmol／L. Compared with the blank control group and NC group， the apoptotic rate increased and the healing rate decreased in the si-YKL-40 group， while the expression of YKL-40， MDR1 and Bcl-2 decreased， while the expression of Bax and caspase-3 increased （P＜0.05）. Conclusion Silencing YKL-40 can significantly increase the sensitivity of Ishikawa／DDP cell lines to cisplatin and promote cell apoptosis， which may be related to down-regulation of MDR1， Bcl-2 expression and up-regulation of Bax and caspase-3 expression.

Key words: Endometrial carcinoma, YKL-40, Cisplatin, Drug resistance

中图分类号:

R737.33

程春来, 丁雯, 车元.

YKL-40调控子宫内膜癌顺铂化疗耐药性的实验研究 [J]. 临床肿瘤学杂志, 2019, 24(2): 113-118.

CHENG Chunlai, DING Wen, CHE Yuan.. Experimental study of YKL-40 regulating cisplatin resistance in endometrial cancer[J]. Chinese Clinical Oncology, 2019, 24(2): 113-118.

参考文献

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

本文评价

推荐阅读 10

[1]	高亚杰¹，关小倩¹，杨海林¹，张　阳²，欧阳学农³，杨建伟⁴，陈　焰⁵,徐建明⁶，赵宣良⁷，王宝成⁸，刘文超⁹，张贺龙¹⁰，南克俊¹¹，王湘辉¹² . 注射用左亚叶酸钙联合治疗晚期胃癌和结直肠癌的Ⅱ期临床研究[J]. 临床肿瘤学杂志, 2009, 14(1): 47 .
[2]	白秀丽¹，马振刚¹，李际君². 紫杉醇联合卡培他滨治疗进展期胃癌的临床观察[J]. 临床肿瘤学杂志, 2009, 14(1): 68 .
[3]	张艳玲，邹　岚，肖　红，黄海辉，谭崇富，阮志华，王　希，梁后杰，庞学利. 影像引导调强放射治疗鼻咽癌[J]. 临床肿瘤学杂志, 2009, 14(1): 51 .
[4]	李　进，曹　君. 大肠癌单克隆抗体治疗新进展[J]. 临床肿瘤学杂志, 2009, 14(1): 1 .
[5]	鲍永仪¹，关乃富¹，程羽青¹. 小细胞肺癌组织中柯萨奇-腺病毒受体阳性表达的意义[J]. 临床肿瘤学杂志, 2009, 14(2): 176 .
[6]	韩　宇^1，2，王　岩¹，徐建明¹，刘烈军¹，赵传华¹，李志强¹，刘建芝¹，邱　卉¹. 诱导化疗后吉非替尼维持治疗晚期非小细胞肺癌的临床研究[J]. 临床肿瘤学杂志, 2009, 14(2): 118 .
[7]	朱　川¹，李　刚¹，任必勇¹，刘必宽¹，邓　超¹，张　军¹，张　力¹，刘学芬¹，熊德明¹. 紫杉醇为主联合化疗方案治疗晚期头颈部肿瘤[J]. 临床肿瘤学杂志, 2009, 14(2): 166 .
[8]	张百红¹，王湘辉¹，凌昌全². ＣＩＳ分期系统对预测肝癌预后的价值研究[J]. 临床肿瘤学杂志, 2009, 14(2): 162 .
[9]	单慧敏¹，陈　琳¹，杨其六¹. 甲状腺微小乳头状癌的临床病理分析[J]. 临床肿瘤学杂志, 2009, 14(2): 170 .
[10]	俞　晶¹，黄云超²，张丽娟³，卢玉波¹. ＨＩＦ-１α和ＧＬＵＴ-１在卵巢癌中的表达及临床意义[J]. 临床肿瘤学杂志, 2009, 14(3): 207 .