| 朱博文,虞海惠,罗 静,韩 笑,陈 慧,邵 荣.分泌蛋白YKL-40增强肿瘤细胞的上皮间质样转化[J].,2022,(2):201-206 |
| 分泌蛋白YKL-40增强肿瘤细胞的上皮间质样转化 |
| Secreted Protein YKL-40 Enhances Epithelial Mesenchymal Transition of Tumor Cells in Vitro |
| 投稿时间：2021-04-28 修订日期：2021-05-23 |
| DOI：10.13241/j.cnki.pmb.2022.02.001 |
| 中文关键词: YKL-40 HCT-116 MDA-MB-231 上皮间质转化 肿瘤进展 |
| 英文关键词: YKL-40 HCT-116 MDA-MB-231 Epithelial mesenchymal transition Tumor progression |
| 基金项目:国家自然科学基金面上项目（81772512） |
|
| 摘要点击次数: 747 |
| 全文下载次数: 411 |
| 中文摘要: |
| 摘要 目的：分泌糖蛋白YKL-40在多种晚期肿瘤病人的血液中显著升高，提示YKL-40蛋白的血浓度是肿瘤恶变的生物标志物。本课题研究YKL-40重组蛋白和过表达YKL-40肿瘤细胞对肿瘤细胞的上皮间质样转化的作用。方法：构建YKL-40过表达的纤维状乳腺癌细胞系MDA-MB-231和非纤维状结肠癌细胞系HCT-116，观察细胞形态学变化，收集细胞和细胞培养液用于Western Blot（WB) 检测YKL-40和上皮间质转化标记蛋白Vimentin和N-cadherin。观察重组蛋白YKL-40对原代MDA-MB-231细胞在无血清条件下的细胞存活影响；另外，用细胞存活试剂盒检测YKL-40过表达HCT-116细胞在无血清的培养液中细胞存活情况。最后，用细胞侵袭试验检测YKL-40过表达MDA-MB-231细胞的侵袭力，并用WB和Zymography来测定细胞分泌MMP9蛋白的表达和酶活性。结果：YKL-40过表达增强MDA-MB-231细胞的形态向上皮间质样转化，并显著提高Vimentin、N-cadherin蛋白的表达，但对HCT-116细胞无法诱导上皮间质样转化。在无血清培养基培养条件下，YKL-40可以增强两种细胞的存活能力，并且YKL-40过表达的MDA-MB-231细胞增强了细胞的侵袭能力，促进了MMP9蛋白表达和蛋白活性。结论：YKL-40可以增加肿瘤细胞的存活力，增强纤维状细胞向上皮间质样转化；并且，YKL-40增加MMP9蛋白表达和活性，增强细胞侵袭力。YKL-40是间质样肿瘤细胞EMT的增强子，此发现为抑制肿瘤恶变提供新靶点。 |
| 英文摘要: |
| ABSTRACT Objective: A large body of evidence has demonstrated that serum levels of secreted protein YKL-40 (CHI3L1) were elevated in a broad type of human advanced cancers, suggesting that serum levels of YKL-40 serve as a cancer biomarker for the malignancy. The current study is to investigate the effects of recombinant protein YKL-40 and YKL-40 over-expressed by tumor cells on tumor epithelial mesenchymal transition (EMT). Methods: Two cancer lines breast cancer cell line MDA-MB-231 that displays a fibroblastic phenotype and colon cancer cell line HCT-116 that does not exhibit the fibroblastic morphology were selected to enforce expression of ectopic YKL-40. After both cell lines were established to stably express YKL-40, the cell phenotype was observed. EMT markers vimentin and N-cadherin proteins were analyzed by Western Blot. The cell cultured media were collected to test YKL-40 levels. Control and HCT-116 cells expressing ectopic YKL-40 were used to test cell survival in the presence of serum free medium for 72 h via a live and dead kit. Concomitantly, recombinant protein YKL-40 was added to parental MDA-MB-231 cells to observe cell survival in the serum-free medium for 72h. MDA-MB-231 cells expressing YKL-40 were employed to examine cell invasion and cultured media were exploited to test MMP9 expression via WB and enzymatic activity via zymography. Results: MDA-MB-231 cells expressing YKL-40 displayed a more mesenchymal phenotype than control cells; whereas HCT-116 cells expressing YKL-40 showed subtle induction of a fibroblastic phenotype. MDA-MB-231 cells expressing YKL-40, but not HCT-116 cells expressing YKL-40, revealed elevated expression of vimentin and N-cadherin. YKL-40 or YKL-40-expressing tumor cells induced tumor cell survival. YKL-40-expressing MDA-MB-231 cells increased invasion and the expression and enzymatic activity of MMP9. Conclusion: YKL-40 has the ability to increase tumor cell survival and enhance EMT, pointing to YKL-40 as a therapeutic target for cancer treatment. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
