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Quote : 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英文版,2025,45(10):1861-1868.[Click to copy ]
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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
Received:March 15, 2025  
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)。
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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