中国癌症杂志
中國癌癥雜誌
중국암증잡지
CHINA ONCOLOGY
2013年
12期
967-973
,共7页
邓佑兴%史惠蓉%李霞%张瑞涛
鄧祐興%史惠蓉%李霞%張瑞濤
산우흥%사혜용%리하%장서도
结肠癌转移相关基因1%卵巢癌%顺铂%细胞外信号激酶
結腸癌轉移相關基因1%卵巢癌%順鉑%細胞外信號激酶
결장암전이상관기인1%란소암%순박%세포외신호격매
Metastasis-associated in colon cancer-1%Ovarian cancer%Cisplatin%Extracellular signal-regulated kinase
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 47.501/47.089μmol/L， P<0.05)，p-ERK1/2蛋白表达下降(0.3979 vs 0.6712/0.6681，P<0.05)，顺铂处理前、后凋亡率增加(1.32%vs 0.66%/0.48%，P<0.05；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。
揹景與目的：有研究髮現，結腸癌轉移相關基因1(metastasis-associated in colon cancer-1，MACC1)在多種腫瘤中高錶達，其通過調節細胞外信號激酶(extracellular signal-regulated kinase，ERK)1/2信號通路影響腫瘤細胞的增殖、凋亡等過程。而對于MACC1在卵巢癌中的作用研究甚少。本研究應用RNA榦擾(RNAi)技術逆轉卵巢上皮性癌耐藥細胞中MACC1基因的錶達，併探討MACC1基因錶達與卵巢癌細胞順鉑耐藥的關繫。方法：前期設計的針對MACC1 mRNA的小髮卡狀RNA(shRNA)轉染卵巢癌順鉑耐藥細胞繫SKOV3/DDP細胞，RT-PCR及Western blot檢測MACC1 mRNA和蛋白錶達的變化；四甲基偶氮唑藍法檢測轉染前後SKOV3/DDP細胞的增殖及對順鉑的半數抑製濃度(IC50)，流式細胞儀測定順鉑處理前後各組細胞凋亡率。Western blot檢測ERK1/2及p-ERK1/2蛋白的變化。實驗分3組：空白組為未經處理的SKOV3/DDP細胞，陰性對照組轉染空質粒p-super-EGFP-1，實驗組轉染重組質粒p-super-EGFP-MACC1 shRNA。結果：與其他兩組細胞相比，實驗組細胞MACC1 mRNA及蛋白降低，對順鉑的IC50較低(26.094 vs 47.501/47.089μmol/L， P<0.05)，p-ERK1/2蛋白錶達下降(0.3979 vs 0.6712/0.6681，P<0.05)，順鉑處理前、後凋亡率增加(1.32%vs 0.66%/0.48%，P<0.05；36.70% vs 18.53%/16.60%，P=0.000)。結論：MACC1基因錶達下調能在一定程度上恢複SKOV3/DDP細胞對順鉑的敏感性，可能與ERK1/2通路有關。
배경여목적：유연구발현，결장암전이상관기인1(metastasis-associated in colon cancer-1，MACC1)재다충종류중고표체，기통과조절세포외신호격매(extracellular signal-regulated kinase，ERK)1/2신호통로영향종류세포적증식、조망등과정。이대우MACC1재란소암중적작용연구심소。본연구응용RNA간우(RNAi)기술역전란소상피성암내약세포중MACC1기인적표체，병탐토MACC1기인표체여란소암세포순박내약적관계。방법：전기설계적침대MACC1 mRNA적소발잡상RNA(shRNA)전염란소암순박내약세포계SKOV3/DDP세포，RT-PCR급Western blot검측MACC1 mRNA화단백표체적변화；사갑기우담서람법검측전염전후SKOV3/DDP세포적증식급대순박적반수억제농도(IC50)，류식세포의측정순박처리전후각조세포조망솔。Western blot검측ERK1/2급p-ERK1/2단백적변화。실험분3조：공백조위미경처리적SKOV3/DDP세포，음성대조조전염공질립p-super-EGFP-1，실험조전염중조질립p-super-EGFP-MACC1 shRNA。결과：여기타량조세포상비，실험조세포MACC1 mRNA급단백강저，대순박적IC50교저(26.094 vs 47.501/47.089μmol/L， P<0.05)，p-ERK1/2단백표체하강(0.3979 vs 0.6712/0.6681，P<0.05)，순박처리전、후조망솔증가(1.32%vs 0.66%/0.48%，P<0.05；36.70% vs 18.53%/16.60%，P=0.000)。결론：MACC1기인표체하조능재일정정도상회복SKOV3/DDP세포대순박적민감성，가능여ERK1/2통로유관。
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 lfow 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 47.501/47.089μmol/L, P<0.05). There was a lower expression of p-ERK1/2 in experimental group (0.3979 vs 00.6712/0.6681, P<0.05). Apoptosis rate was significantly higher in the experimental group before and after treatment of cisplatin (1.32%vs 0.66%/0.48%, P<0.05;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.
