| 仇建玲,吴胜文,俞进友,等.胶原蛋白 Ⅴ型 2链对胃癌细胞增殖、迁移和侵袭及肿瘤生长的调控作用[J].安徽医药,2024,28(2):280-285. |
| 胶原蛋白 Ⅴ型 2链对胃癌细胞增殖、迁移和侵袭及肿瘤生长的调控作用 |
| Experimental study on col5a2 regulating proliferation, migration, invasion and tumor growth of gastric cancer cells |
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| DOI：10.3969/j.issn.1009-6469.2024.02.015 |
| 中文关键词: 胃肿瘤 胶原蛋白 Ⅴ型 2链 裸鼠成瘤 增殖 迁移 侵袭 |
| 英文关键词: Stomach neoplasms Collagen type V 2 chain Tumorigenesis in nude mice Proliferation Migration Invasion |
| 基金项目:江苏省卫生健康委医学科研立项项目( Z2021027) |
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| 中文摘要: |
| 目的探究胶原蛋白 Ⅴ型 2链( COL5A2)对胃癌细胞增殖、迁移和侵袭及肿瘤生长的调控作用。方法于 2022年 1―3月开展研究，基于 TCGA数据库分析 COL5A2在胃癌组织、正常组织中的表达差异， COL5A2对胃癌分期的作用及对病人预后的价值。荧光定量聚合酶链式反应( qRT-PCR)、蛋白质印迹法( WB)检测正常人胃黏膜上皮细胞 GES-1、胃癌细胞 MGC-803、 MKN-45、SGC7901中 COL5A2的表达。慢病毒介导稳定转染 shRNA-COL5A2、shRNA、NC的 MKN-45细胞。克隆形成实验、细胞计数试剂盒( CCK8)、 5-乙炔基 -2'-脱氧尿苷( EdU)染色检测细胞的增殖；碘化丙啶( PI)染色检测细胞周期分布； Transwell实验、伤口愈合实验检测细胞的迁移、侵袭能力；裸鼠成瘤实验检测肿瘤的生长；免疫组织化学实验检测肿瘤组织 PCNA蛋白。结果胃癌组织 COL5A2相对表达量为 4.74±0.45，显著高于正常组织的 2.53±0.26(P<0.05)与正常组织相比，胃癌组织中 COL5A2升高，与病人临床分期、预后差有关。 GES-1组 COL5A2相对表达量为 0.25±0.04，显著，低于胃癌细胞中 MGC-803组0.47±0.05、MKN-45组 0.72±0.08、SGC7901组 0.54±0.04(P<0.05)，其中 MKN-45升高幅度最大。与 shRNA组细胞相比， shRNA-COL5A2组细胞 COL5A2表达降低，细胞的克隆形成数、在 48、72 h的活性、 EdU阳性率、迁移和侵袭数量、划痕愈合率均降低，细胞周期阻滞 G1/S期， CyclinD1和 CDK4表达降低( P<0.05)。异种移植裸鼠成瘤的体积、质量均降低，肿瘤 PCNA表达也降低。结论 COL5A2在胃癌中高表达，下调 COL5A2抑制癌细胞增殖、迁移侵袭及肿瘤的生长。 |
| 英文摘要: |
| Objective To investigate the regulation of collagen type V 2 chain (COL5A2) on the proliferation, migration, invasion andtumor growth of gastric cancer cells.Methods To start the study from January to March 2022, Based on TCGA database, the expres.sion difference of COL5A2 in gastric cancer and normal tissues, the effect of COL5A2 on gastric cancer staging and the value of progno.sis were analyzed. The expression of COL5A2 in normal human gastric epithelial cells GES-1, gastric cancer cells MGC-803, MKN-45 and SGC7901 was detected by fluorescence quantitative polymerase chain reaction (qRT PCR) and Western blotting (WB). MKN-45 cells stably transfected with shrna-COL5A2, shRNA and NC mediated by lentivirus. Cell proliferation was detected by clone formationtest, cell counting Kit (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) staining; Propidium iodide (PI) staining was used to detect the dis.tribution of cell cycle; Transwell test and wound healing test were used to detect the migration and invasion ability of cells; The growthof tumor was detected by tumorigenesis experiment in nude mice; PCNA protein in tumor tissue was detected by immunohistochemistry.Results The relative expression of COL5A2 in gastric cancer tissues was (4.74±0.45), which was significantly higher than that in nor.mal tissues (2.53±0.26) (P<0.05),compared with normal tissues, COL5A2 in gastric cancer increased, which was related to the clinicalstage and poor prognosis. The relative expression of COL5A2 in GES-1 group was (0.25±0.04), which was significantly lower than that in MGC-803 group (0.47±0.05), MKN-45 group (0.72±0.08) and SGC7901 group (0.54±0.04) (P<0.05), and the increase of MKN-45 was the largest. Compared with shRNA group, the expression of COL5A2 in shrna-COL5A2 group decreased, the number of clone for.mation, activity at 48 and 72 hours, EdU positive rate / migration and invasion number, scratch healing rate, cell cycle arrest G1/Sphase, and the expression of CyclinD1 and CDK4 decreased (P<0.05). The volume and mass of xenografted nude mice decreased, and the expression of PCNA also decreased.Conclusion COL5A2 is highly expressed in gastric cancer, which promotes the proliferation,migration and invasion of cancer cells and contributes to the growth of tumor. |
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