| 引用本文: |
赵沅, 黄启宣, 胡响当.天马颗粒Ⅲ含药血清通过下调HK2表达抑制结直肠癌细胞迁移和侵袭[J].湖南中医药大学学报,2025,45(10):1861-1868[点击复制] |
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| 天马颗粒Ⅲ含药血清通过下调HK2表达抑制结直肠癌细胞迁移和侵袭 |
| 赵沅,黄启宣,胡响当 |
| (湖南中医药大学第二附属医院, 湖南 长沙 410005;曲靖市中医医院, 云南 曲靖 655099) |
| 摘要: |
| 目的 探讨天马颗粒Ⅲ含药血清（TMKL）对结直肠癌（CRC）细胞侵袭、迁移的影响，验证天马颗粒Ⅲ含药血清是否通过调控己糖激酶2（HK2）的表达从而抑制CRC细胞的侵袭、迁移。方法 以结直肠癌SW480细胞作为研究对象，采用CCK-8法筛选TMKL作用SW480的最佳浓度，将SW480细胞分为空白血清组（BS组）、TMKL含药血清组（TMKL组），以验证TMKL对SW480细胞的作用。通过质粒转染技术构建HK2敲低SW480细胞株后，将细胞随机分为空白质粒组（sh-NC组）、敲低HK2组（sh-HK2组）、敲低HK2后使用空白血清干预组（sh-HK2+BS组）、敲低HK2后使用TMKL含药血清干预组（sh-HK2+TMKL组）。通过CCK-8法检测细胞增殖能力；Transwell法检测细胞侵袭、迁移能力；RT-qPCR法、Western blot法检测细胞HK2、上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）的mRNA及蛋白表达水平。结果 与BS组相比，TMKL组细胞增殖、侵袭、迁移能力明显下降（P＜0.01），HK2的mRNA及蛋白表达水平明显降低（P＜0.01），E-cadherin表达水平明显升高（P＜0.01），N-cadherin 表达水平明显降低（P＜0.01）。在HK2敲低背景下，与sh-NC组相比，sh-HK2组细胞增殖、侵袭和迁移能力显著下降（P＜0.01），E-cadherin的mRNA及蛋白表达水平显著升高（均P＜0.01），HK2和N-cadherin的mRNA及蛋白水平显著降低（P＜0.01）。与sh-HK2组相比，sh-HK2+TMKL组HK2的mRNA及蛋白表达水平显著降低（P＜0.01）。结论 TMKL含药血清可抑制结直肠癌细胞增殖、侵袭和迁移，该作用机制可能与下调HK2表达，从而抑制细胞EMT进程有关。 |
| 关键词: 结直肠癌 天马颗粒Ⅲ 己糖激酶2 细胞增殖 细胞迁移 细胞侵袭 |
| DOI：10.3969/j.issn.1674-070X.2025.10.008 |
| 投稿时间：2025-03-15 |
| 基金项目:湖南省卫生健康委员会课题（C202304017114）；湖南省中医药管理局科研项目（D2023008）；湖南中医药大学校级科研项目（2021XJJJ054）。 |
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| Tianma Granules Ⅲ-containing serum inhibits migration and invasion of colorectal cancer cells by downregulating HK2 expression |
| ZHAO Yuan, HUANG Qixuan, HU Xiangdang |
| (The Second Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410005, China;Qujing Hospital of Traditional Chinese Medicine, Qujing, Yunnan 655099, China) |
| Abstract: |
| Objective To investigate the effects of Tianma Granules Ⅲ (TMKL)-containing serum on the invasion and migration of colorectal cancer (CRC) cells, and to verify whether TMKL inhibits these processes by regulating hexokinase 2 (HK2) expression. Methods Human CRC SW480 cells were used as the research subject. The optimal concentration of TMKL for treating SW480 cells was first determined using the CCK-8 assay. SW480 cells were divided into blank serum group (BS group) and TMKL-containing serum group (TMKL group) to verify the effects of TMKL. Subsequently, a HK2-knockdown SW480 cell line was established via plasmid transfection, and these cells were randomly assigned into blank plasmid group (sh-NC group), HK2-knockdown group (sh-HK2 group), HK2-knockdown+blank serum intervention group (sh-HK2+BS group), and HK2-knockdown+TMKL-containing serum intervention group (sh-HK2+TMKL group). Cell proliferation was evaluated by CCK-8 assay; cell invasive and migratory capacities were assessed using Transwell assay; mRNA and protein expression levels of HK2, E-cadherin, and N-cadherin were measured by RT-qPCR and Western blot, respectively. Results Compared with the BS group, the TMKL group exhibited significantly reduced capacities of cell proliferation, invasion, and migration (all P＜0.01), along with markedly lower HK2 mRNA and protein expression levels (P＜0.01), obviously upregulated E-cadherin expression (P＜0.01), and significantly downregulated N-cadherin expression (P＜0.01). Under the condition of HK2 knockdown, compared with the sh-NC group, the sh-HK2 group exhibited significantly reduced capacities of cell proliferation, invasion, and migration (all P＜0.01), significantly upregulated mRNA and protein expression of E-cadherin (P＜0.01), and significantly downregulated mRNA and protein expression of HK2 and N-cadherin (P＜0.01). Compared with the sh-HK2 group, the sh-HK2+TMKL group showed significantly lower mRNA and protein expression levels of HK2 (P＜0.01). Conclusion TMKL-containing serum can inhibit the proliferation, invasion, and migration of CRC cells, and the mechanism may be related to the downregulation of HK2 expression, thereby inhibiting the process of epithelial-mesenchymal transition (EMT). |
| Key words: colorectal cancer Tianma Granules Ⅲ hexokinase 2 cell proliferation cell migration cell invasion |
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