| 雷建卫,汪媛,贺利荣.分泌型卷曲相关蛋白 1通过 MAPK/ERK信号通路抑制结直肠癌细胞的迁移侵袭[J].安徽医药,2022,26(4):777-782. |
| 分泌型卷曲相关蛋白 1通过 MAPK/ERK信号通路抑制结直肠癌细胞的迁移侵袭 |
| SFRP1 inhibits the migration and invasion of colorectal cancer cells through MAPK/ERK signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2022.04.031 |
| 中文关键词: 结直肠肿瘤 细胞外信号调节 MAP激酶类 钙黏着糖蛋白类 波形蛋白 分泌型卷曲相关蛋白 1 MAPK/ ERK信号通路 迁移 侵袭 |
| 英文关键词: Colorectal neoplasms Extracellular signal-regulated MAP kinases Cadherins Vimentin SFRP1 MAPK/ERK signaling pathway Migration Invasion |
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| 中文摘要: |
| 目的探讨分泌型卷曲相关蛋白 1(SFRP1)对结直肠癌细胞迁移和侵袭的影响机制。方法本研究起止时间 2018年 11月至 2019年 6月。运用实时荧光定量反转录聚合酶链反应( qRT-PCR)检测正常结肠上皮细胞 HCoEpiC、人结直肠癌细胞 HCT8、SW480、RKO中 SFRP1的 mRNA表达；将 pcDNA 3.1组(转染 pcDNA 3.1)pcDNA 3.1-SFRP1组(转染 pcDNA 3.1-SFRP1)、 pcDNA 3.1-SFRP1+DMSO组［转染 pcDNA 3.1-SFRP1，再用二甲基亚砜( DMSO)处、理］、 pcDNA 3.1-SFRP1+IGF组［转染 pcDNA 3.1-SFRP1，再用丝裂原活化蛋白激酶( MAPK)/细胞外信号调节激酶( ERK)信号通路激活剂( IGF1)处理］均用脂质体法转染至 HCT8细胞。蛋白质印迹法( Western blotting)检测细胞中 SFRP1、波形蛋白(Vimentin)、纤维黏连蛋白(FN)、，基质金属蛋白酶 9(MMP-9)、 N-钙黏蛋白( N-cadherin)、上皮钙黏蛋白( E-cadherin)、 MAPK/ERK信号通路关键基因( Ras)、雷帕霉素靶蛋白(mTOR)、细胞外信号调节激酶 1/2(ERK1/2)、磷酸化细胞外信号调节激酶 1/2(p-ERK1/2)、细胞外信号调节激酶( ERK)的蛋白表达；迁徙实验( Transwell)检测细胞的迁移侵袭。结果与正常结肠上皮细胞 HCoEpiC相比，人结直肠癌细胞 HCT8、SW480、 RKO中 SFRP1的表达显著降低［mRNA(1.00±0.06)比( 0.36±0.03)、(0.62±0.05)、(0.59±0.05)；蛋白( 1.00±0.04)比( 0.24±0.02)、(0.48±0.04)、(0.47±0.04)均 P<0.05］。过表达 SFRP1可抑制 HCT8细胞的迁移( 120±11)比( 65±6)和侵袭( 98±8)比( 47±4)并下调Vimentin(0.96±0.05)比，(0.23±0.02)、FN(1.00±0.07)比(0.51±0.04)、MMP-9(0.98±0.08)比(0.35±0.03)、N-cadherin(1.01±0.09)比(0.43±0.04)上调 E-cadherin(0.99±0.06)比( 3.71±0.27)抑制 MAPK/ERK信号通路关键蛋白 Ras(0.97±0.07)比( 0.34±0.03)、mTOR(1.03±0.07)比(0.42±0.04)、 p-ERK1/2(0.98±0.08)比(0.29±0.03)和 ERK(1.01±0.06)比( 0.31±0.03)的表达。激活 MAPK/ERK信号通路可逆转过表达 SFRP1对结直肠癌细胞迁移和侵袭的抑制作用。结论 SFRP1可抑制结直肠癌细胞的迁移和侵袭，其机制可能与失活 MAPK/ERK信号通路相关，将可为 SFRP1用于结直肠癌的治疗提供依据。 |
| 英文摘要: |
| Objective To investigate the mechanism of secretory crimp protein 1 (SFRP1) on migration and invasion of colorectal cancer cells.Methods The start and end time of study was from November 2018 to June 2019. Real-time fluorescent quantitative re verse transcription polymerase chain reaction(qRT-PCR) was used to detect the expression of SFRP1 mRNA in normal colonic epitheli al cells HCoEpiC, human colorectal cancer cells HCT8, SW480 and RKO. pcDNA 3.1 group (transfected pcDNA 3.1), pcDNA 3.1SFRP1 group (transfected pcDNA 3.1-SFRP1), pcDNA 3.1-SFRP1+DMSO [transfected pcDNA3.1-SFRP1 and treated with dimethyl sulfoxide (DMSO)], pcDNA 3.1-SFRP1+IGF (transfected pcDNA 3.1-SFRP1 and treated with MAPK/ERK signaling activator IGF1), allwere transfected into HCT8 cells by liposome method. Western blotting (Western blot) was used to detect the proteins expression ofSFRP1, Vimentin (Vimentin), fibronectin (FN), matrix metalloproteinase 9 (MMP-9), N-cadherin (N-cadherin) and epithelial cadherin (E-cadherin), key genes of mitogen activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) signaling path way (Ras), rapamycin target protein (mTOR), extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2), extracellular signal-regulated kinase (ERK). The cell migration and invasion were detected by Tran swell chamber.Results Compared with normal colonic epithelial cells HCoEpiC, the expression of SFRP1 in human colorectal cancercells HCT8, SW480 and RKO were significantly decreased [mRNA (1.00±0.06) vs. 0.36±0.03),(0.62±0.05,(0.59±0.05); protein (1.00±0.04)vs.(0.24±0.02),(0.48±0.04),(0.47±0.04)] (P<0.05). Overexpression SFRP1 inhibited the migration (120±11)vs.(65±6) and invasion (98±8) vs. (47±4), down-regulated vimentin (0.96±0.05) vs. (0.23±0.02), FN (1.00±0.07)vs. (0.51±0.04), MMP-9 (0.98±0.08)vs.(0.35± 0.03), N-cadherin (1.01±0.09) vs.(0.43±0.04), up-regulated E-cadherin (0.99±0.06)vs.(3.71±0.27), and inhibited the expression of key proteins of MAPK/ERK signaling pathway including Ras (0.97±0.07)vs. (0.34±0.03), mTOR (1.03±0.07)vs. (0.42±0.04), p-ERK1/2 (0.98±0.08)vs. (0.29±0.03) and ERK (1.01±0.06)vs. (0.31±0.03). Activation MAPK/ERK signaling pathway reversed the inhibitory ef fect of overexpression SFRP1 on migration and invasion of colorectal cancer cells.Conclusions SFRP1 can inhibit the migration andinvasion of colorectal cancer cells, and its mechanism may be related to the inactivation of MAPK/ERK signaling pathway, which willprovide a basis for the treatment of colorectal cancer with SFRP1. |
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