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朱思洵, 姚昶, 杨静, 苏彬, 赵倩馨, 蒋立新.基于APLN/PI3K/Akt信号通路探讨柴胡疏肝散加减对三阴性乳腺癌细胞增殖、迁移与侵袭的影响[J].湖南中医药大学学报英文版,2025,45(2):228-238.[Click to copy
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| 基于APLN/PI3K/Akt信号通路探讨柴胡疏肝散加减对三阴性乳腺癌细胞增殖、迁移与侵袭的影响 |
| 朱思洵,姚昶,杨静,苏彬,赵倩馨,蒋立新 |
| (江阴市中医院 乳腺外科, 江苏 无锡 214400;江苏省中医院 乳腺外科, 江苏 南京 210000) |
| 摘要: |
| 目的 基于转录组学测序探究柴胡疏肝散加减含药血清通过爱帕琳肽（APLN）/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路对三阴性乳腺癌MDA-MB-231细胞增殖、迁移与侵袭的影响。方法 采用SD大鼠制备柴胡疏肝散加减含药血清和对照血清。不同比例对照血清和含药血清干预MDA-MB-231细胞24 h，通过CCK-8法检测细胞增殖活力，筛选最佳比例；采用转录组测序分析对照血清组和含药血清组细胞中差异表达基因和信号通路。将APLN siRNA或过表达质粒转染细胞，以敲低或过表达APLN；采用CCK-8法、划痕与Transwell侵袭实验分别检测含药血清联合APLN敲低或过表达干预后细胞的增殖、迁移与侵袭能力；采用RT-PCR以及Western blot法分别检测APLN、PI3K、Akt基因与蛋白的表达情况。结果 与对照血清组比较，含药血清组中细胞的增殖活力、迁移与侵袭能力降低（P<0.05）。转录组结果显示，相对于对照血清组，含药血清组中上调116个基因，下调66个基因；这些差异基因主要富集于PI3K/Akt、丝裂原活化蛋白激酶（MAPK）等肿瘤相关的信号通路。在Top30差异基因中，含药血清降低APLN基因和蛋白的表达（P<0.05），并抑制APLN下游PI3K/Akt信号通路的激活（P<0.05）；APLN敲低进一步增强含药血清对细胞增殖、迁移、侵袭以及PI3K/Akt通路激活的抑制作用（P<0.05），而APLN过表达则削弱了含药血清的上述作用（P<0.05）。结论 柴胡疏肝散加减含药血清可通过负调控APLN的表达拮抗PI3K/Akt信号通路的激活来抑制三阴性乳腺癌细胞增殖、迁移与侵袭，以达到抗癌的疗效。 |
| 关键词: 柴胡疏肝散加减 三阴性乳腺癌 APLN/PI3K/Akt信号通路 增殖 迁移 侵袭 |
| DOI：10.3969/j.issn.1674-070X.2025.02.006 |
| Received:September 19, 2024 |
| 基金项目:无锡市卫生健康委员会重点项目（Z202203）。 |
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| Effects of modified Chaihu Shugan Powder on proliferation, migration, and invasion of triple-negative breast cancer cells based on APLN/PI3K/Akt signaling pathway |
| ZHU Sixun, YAO Chang, YANG Jing, SU Bin, ZHAO Qianxin, JIANG Lixin |
| (Department of Breast Surgery, Jiangyin Hospital of Chinese Medicine, Wuxi, Jiangsu 214400, China;Department of Breast Surgery, Jiangsu Province Hospital of Chinese Medicine, Nanjing, Jiangsu 210000, China) |
| Abstract: |
| Objective To explore the effects of modified Chaihu Shugan Powder medicated serum on proliferation, migration, and invasion of triple-negative breast cancer MDA-MB-231 cells through the APLN/PI3K/Akt signaling pathway based on transcriptome sequencing. Methods SD rats were used to prepare modified Chaihu Shugan Powder medicated serum and control serum. MDA-MB-231 cells were treated with different proportions of control and medicated serums for 24 h. Cellular proliferation activity was assessed using the CCK-8 assay to determine the optimal proportion. Transcriptome sequencing was employed to analyze differentially expressed genes and signaling pathways in cells from the control serum group and the medicated serum group. Cells were transfected with APLN siRNA or overexpression plasmid to knock down or overexpress APLN. The CCK-8 assay, wound healing assay, and Transwell invasion assay were used to check cellular proliferation, migration, and invasion abilities, respectively, after intervention with the medicated serum combined with APLN knockdown or overexpression. RT-PCR and Western blot were utilized to determine the expressions of APLN, PI3K, and Akt genes and proteins. Results Compared with the control serum group, the cellular proliferation activity, migration, and invasion abilities in the medicated serum group decreased (P<0.05). Transcriptomic results showed that, compared with the control serum group, 116 genes were significantly upregulated and 66 genes were downregulated in the medicated serum group. These differential genes were primarily enriched in tumor-related signaling pathways such as PI3K/Akt and mitogen-activated protein kinase (MAPK). Among the top 30 differential genes, the medicated serum reduced the expressions of APLN gene and protein (P<0.05) and inhibited the activation of the APLN downstream PI3K/Akt signaling pathway (P<0.05). APLN knockdown further enhanced the inhibitory effects of the medicated serum on cell proliferation, migration, invasion, and PI3K/Akt pathway activation (P<0.05), whereas APLN overexpression attenuated these effects of the medicated serum (P<0.05). Conclusion The modified Chaihu Shugan Powder medicated serum can inhibit the proliferation, migration, and invasion of triple-negative breast cancer cells by negatively regulating APLN expression, thereby antagonizing the activation of PI3K/Akt signaling pathway and exerting anti-cancer effects. |
| Key words: modified Chaihu Shugan Powder medicated serum triple-negative breast cancer APLN/PI3K/Akt signaling pathway proliferation migration invasion |
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