| 郑贺予,孔庆权,毛美琪,等.肌切蛋白可变剪接在神经胶质瘤中的作用及机制研究[J].肿瘤学杂志,2026,32(10):866-875. |
| 肌切蛋白可变剪接在神经胶质瘤中的作用及机制研究 |
| The Role and Mechanism of Scinderin Alternative Splicing in Glioma |
| 投稿时间：2025-05-22 |
| DOI：10.11735/j.issn.1671-170X.2026.10.B008 |
|
 |
| 中文关键词: 神经胶质瘤 肌切蛋白 可变剪接 恶性生物学行为 p53 |
| 英文关键词:glioma scinderin alternative splicing malignant biological behavior p53 |
| 基金项目:辽宁省教育厅科学技术研究项目（JYTMS20231388）；辽宁省科技计划联合计划（自然科学基金-面上项目）（2024-MSLH-458）；辽宁省科学技术计划（面上项目）（2024-MS-224）；辽宁省科技计划联合计划（技术攻关计划项目）（2024JH2/102600235）；沈阳医学院硕士研究生科技创新基金（Y20240507） |
|
| 摘要点击次数: 0 |
| 全文下载次数: 0 |
| 中文摘要: |
| 摘 要：［目的］ 探究神经胶质瘤中肌切蛋白（scinderin，SCIN）的不同可变剪接形式对肿瘤细胞恶性生物学行为的影响及其相关分子机制。［方法］ 采用14例神经胶质瘤患者肿瘤组织标本及人神经胶质瘤细胞系U118、U251进行实验。采用Western Blot检测神经胶质瘤不同细胞系、不同级别神经胶质瘤组织中SCIN及其可变剪接体的表达情况。运用CRISPR/Cas9技术建立SCIN敲除神经胶质瘤细胞模型，采用Westren Blot检测其转染效率；采用CCK-8、克隆形成法检测SCIN缺失后对人神经胶质瘤细胞增殖及克隆形成能力的影响；采用划痕实验、Transwell方法检测SCIN缺失后对人神经胶质瘤细胞迁移和侵袭的影响。通过质粒转染在基因敲除细胞模型中过表达SCIN不同可变剪接体，探究SCIN不同可变剪接体对神经胶质瘤增殖、迁移、侵袭等恶性生物学行为的影响。采用Western Blot检测SCIN及其可变剪接体与p53的相关性。［结果］ 临床样本检测结果显示，神经胶质瘤组织中SCIN的可变剪接体存在异常表达。CCK-8结果表明，SCIN缺失组24 h细胞增殖率显著低于对照组（U118：89.97%±2.46% vs 116.63%±12.47%，t=3.69，P<0.01；U251：50.61%±3.67% vs 65.33%±11.27%，t=8.48，P<0.01），48 h SCIN缺失组细胞增殖率较对照组进一步显著下降（U118：37.92%±0.80% vs 55.84%±8.44%，t=8.16，P<0.01；U251：38.05%±0.98% vs 59.49%±4.20%，t=14.25，P<0.01），72 h SCIN缺失组细胞增殖率仍显著低于对照组（U118：28.06%±4.75% vs 46.02%±7.72%，t=15.30 ，P<0.01；U251：21.42%±5.35% vs 23.41%±4.02%，t=17.36，P<0.01）。克隆形成结果表明，SCIN缺失抑制了细胞克隆形成率（U118：t=2.34，P<0.05；U251：t=2.85，P<0.05）。划痕实验结果显示，SCIN缺失后细胞迁移能力显著下降（U118：t=9.72，P<0.01；U251：t=19.00，P<0.01）。Transwell结果显示，SCIN缺失削弱了细胞的迁移和侵袭能力，穿膜细胞数均明显低于对照组（迁移：U118：t=10.03，P<0.01；U251：t=6.68，P<0.01；侵袭：U118：t=6.862，P<0.01；U251：t=5.170，P<0.01）。与过表达全长SCIN细胞相比，468aa亚型异构蛋白则发挥了促进作用，能显著增强神经胶质瘤细胞的迁移（U118：F=25.08，P＜0.01；U251：F=16.72，P＜0.01）和侵袭（U118：F=27.93，P＜0.01；U251：F=26.45，P＜0.01）能力。［结论］ 原癌基因SCIN在神经胶质瘤中存在可变剪接形式，其中468aa亚型异构蛋白异常高表达具有“促癌”作用；此外，SCIN的580aa剪接体的形成与p53蛋白的功能异常密切相关。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effects of different alternative splicing isoforms of scinderin (SCIN) on the malignant biological behavior of tumor cells in glioma and their underlying molecular mechanisms. ［Methods］ Tumor tissues from 14 glioma patients and human glioma cell lines (U118 and U251) were analyzed. The expression of SCIN and its alternative splicing isoforms in different glioma cell lines and glioma tissues with different grades was detected by Western blot. CRISPR/Cas9 technology was employed to establish SCIN knockout glioma cell models， and Western blot was used to verify transfection efficiency. The effects of SCIN deletion on the proliferation and colony formation of human glioma cells were assessed by CCK-8 and colony formation assays. The impact of SCIN deletion on the migration and invasion of human glioma cells was evaluated by scratch wound healing and Transwell assays. By overexpressing different SCIN alternative splicing isoforms in the knockout cell models via plasmid transfection， the effects of different SCIN splicing isoforms on glioma proliferation， migration， invasion， and other malignant biological behaviors were explored. Western blot was used to examine the correlation between SCIN and its splicing isoforms with p53. ［Results］ Clinical sample testing results indicated that there was abnormal expression of alternative splicing variants of SCIN in glioma tissues. CCK-8 results showed that the cell proliferation rate at 24 hours in the SCIN deletion group was significantly lower than that in the control group(U118： 89.97%±2.46% vs 116.63%±12.47%， t=3.69， P<0.01； U251： 50.61%±3.67% vs 65.33%±11.27%， t=8.48， P<0.01). At 48 hours， the cell proliferation rate further decreased significantly compared to the control group (U118： 37.92%±0.80% vs 55.84%±8.44%， t=8.16， P<0.01； U251： 38.05%±0.98% vs 59.49%±4.20%， t=14.25， P<0.01) . At 72 hours， the cell proliferation rate remained significantly lower than that in the control group (U118： 28.06%±4.75% vs 46.02%±7.72%， t=15.30， P<0.01； U251： 21.42%±5.35% vs 23.41%±4.02%， t=17.36， P<0.01). Colony formation results indicated that SCIN deletion inhibited cell colony formation rate (U118： t=2.34， P<0.05； U251： t=2.85， P<0.05). Scratch assay results showed that cell migration ability was significantly reduced after SCIN deletion (U118： t=9.72， P<0.01； U251： t=19.00， P<0.01). Transwell results demonstrated that SCIN deletion impaired cell migration and invasion abilities， with the number of transmembrane cells significantly lower than that in the control group (migration： U118： t=10.03， P<0.01； U251： t=6.68， P<0.01； invasion： U118： t=6.862， P<0.01； U251： t=5.170， P<0.01). Reconstitution experiments demonstrated that the 468aa isoform specifically enhanced the migration (U118： F=25.08， P<0.01； U251： F=16.72， P<0.01) and invasion abilities (U118： F=27.93， P<0.01； U251： F=26.45， P<0.01) of glioma cells more potently than the full-length SCIN. ［Conclusion］ The proto-oncogene SCIN exhibits alternative splicing isoforms in glioma， with abnormal high expression of the 468aa isoform protein playing a “cancer-promoting” role. Additionally， the formation of the 580aa splicing isoform of SCIN is closely related to abnormal p53 protein function. |
|
在线阅读
查看全文 查看/发表评论 下载PDF阅读器 |