Effect of MACC1 gene suppression on the proliferation of SKOV3/DDP cells and its chemosensitivity to cisplatin

邓佑兴; 史惠蓉; 李霞; 张瑞涛

doi:10.3969/j.issn.1007-3969.2013.12.006

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Effect of MACC1 gene suppression on the proliferation of SKOV3/DDP cells and its chemosensitivity to cisplatin

更新时间：2025-12-31

- Effect of MACC1 gene suppression on the proliferation of SKOV3/DDP cells and its chemosensitivity to cisplatin
- China Oncology Vol. 23, Issue 12, Pages: 967-973(2013)
- 作者机构：
  
  郑州大学第一附属医院妇产科,河南,郑州,450052
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.1007-3969.2013.12.006
  CLC：
- Published Online：17 February 2014，
  
  Published：17 February 2013
- 稿件说明：
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邓佑兴,史惠蓉,李霞,张瑞涛. MACC1基因干扰对卵巢上皮性癌细胞增殖和顺铂耐药的影响[J]. 中国癌症杂志, 2013, 23(12): 967-973.

邓佑兴, 史惠蓉, 李霞, et al. Effect of MACC1 gene suppression on the proliferation of SKOV3/DDP cells and its chemosensitivity to cisplatin[J]. China Oncology, 2013, 23(12): 967-973.
邓佑兴,史惠蓉,李霞,张瑞涛. MACC1基因干扰对卵巢上皮性癌细胞增殖和顺铂耐药的影响[J]. 中国癌症杂志, 2013, 23(12): 967-973. DOI： 10.3969/j.issn.1007-3969.2013.12.006.

邓佑兴, 史惠蓉, 李霞, et al. Effect of MACC1 gene suppression on the proliferation of SKOV3/DDP cells and its chemosensitivity to cisplatin[J]. China Oncology, 2013, 23(12): 967-973. DOI： 10.3969/j.issn.1007-3969.2013.12.006.

摘要

背景与目的：有研究发现，结肠癌转移相关基因1(metastasis-associated in colon cancer-1，MACC1)在多种肿瘤中高表达，其通过调节细胞外信号激酶(extracellular signal-regulated kinase，ERK)1/2信号通路影响肿瘤细胞的增殖、凋亡等过程。而对于MACC1在卵巢癌中的作用研究甚少。本研究应用RNA干扰(RNAi)技术逆转卵巢上皮性癌耐药细胞中MACC1基因的表达，并探讨MACC1基因表达与卵巢癌细胞顺铂耐药的关系。方法：前期设计的针对MACC1 mRNA的小发卡状RNA(shRNA)转染卵巢癌顺铂耐药细胞系SKOV3/DDP细胞，RT-PCR及Western blot检测MACC1 mRNA和蛋白表达的变化；四甲基偶氮唑蓝法检测转染前后SKOV3/DDP细胞的增殖及对顺铂的半数抑制浓度(IC50)，流式细胞仪测定顺铂处理前后各组细胞凋亡率。Western blot检测ERK1/2及p-ERK1/2蛋白的变化。实验分3组：空白组为未经处理的SKOV3/DDP细胞，阴性对照组转染空质粒p-super-EGFP-1，实验组转染重组质粒p-super-EGFP-MACC1 shRNA。结果：与其他两组细胞相比，实验组细胞MACC1 mRNA及蛋白降低，对顺铂的IC50较低(26.094 vs 47.501/47.089 μmol/L，P0.05)，p-ERK1/2蛋白表达下降(0.3979 vs 0.6712/0.6681，P0.05)，顺铂处理前、后凋亡率增加(1.32% vs 0.66%/0.48%，P0.05；36.70% vs 18.53%/16.60%，P=0.000)。结论：MACC1基因表达下调能在一定程度上恢复SKOV3/DDP细胞对顺铂的敏感性，可能与ERK1/2通路有关。

Abstract

Background and purpose: The metastasis-associated in colon cancer-1 (MACC1) is highly expressed in different cancers and has an effect on the proliferation and apoptosis of tumor cells through the regulation of extracellular signal-regulated kinase (ERK)1/2 pathway. However

the role of MACC1 in ovarian cancer has been rarely studied. The study was aimed to suppress MACC1 gene expression by siRNA and explore the relationship between MACC1 expression and chemosensitivity to cisplatin in ovarian cancer cell line SKOV3/DDP. Methods: Empty plasmid p-super-EGFP-1 (negative control group) and p-super-EGFP-MACC1 shRNA (experimental group) were transfected into ovarian cancer cell SKOV3/DDP respectively. SKOV3/DDP cells without transfection were used as blank group. Then

MACC1 mRNA and protein levels were measured by RT-PCR and Western blot

respectively. Cell proliferation and IC50 of cisplatin was determined by methyl thiazolyl tetrazolium test (MTT). Apoptosis rate was determined by flow cytometer (FCM). ERK1/2 and p-ERK1/2 protein levels were determined by Western blot. Results: Compared with those in blank and negative control groups

MACC1 mRNA and protein levels deceased in experimental group. The IC50 of cisplatin in experimental group was lower than that in the other groups (26.094 vs 47.501/47.089 μmol/L

P0.05). There was a lower expression of p-ERK1/2 in experimental group (0.3979 vs 00.6712/0.6681

P0.05). Apoptosis rate was significantly higher in the experimental group before and after treatment of cisplatin (1.32% vs 0.66%/0.48%

P0.05; 36.70% vs 18.53%/16.60%

P=0.000). Conclusion: MACC1 gene may be involved in cisplatin resistance phenomenon in SKOV3/DDP cells through ERK1/2 pathway.

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