Department of Gastroenterology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, 210029, China.
Cell Mol Biol (Noisy-le-grand). 2020 Jun 5;66(3):119-124.
This study aims to study the effect and mechanism of Patrinia herba aqueous extract on proliferation, apoptosis, invasion and migration of hepatocellular carcinoma cells. Hepatocellular carcinoma cells MHCC97-H were treated with 2.5, 5, 10 mg/mL P. herba aqueous extract, cell counting kit 8 (CCK-8), flow cytometry, plate cloning experiments, and Transwell measured cell survival, apoptosis, colony formation, invasion, and migration, respectively. Real-time quantitative PCR (qPCR) and western blot were used to detect long non-coding RNA (lncRNA) HTR2A-AS1 and expression of proteins P21, Caspase-3, E-cadherin and matrix metalloproteinase-2 (MMP-2), respectively. Transfected pcDNA3.1-HTR2A-AS1 in MHCC97-H cells, or transfected si-HTR2A-AS1 and treat with 10 mg/mL P. herba aqueous extract to evaluate their roles in cell proliferation, apoptosis, invasion, and migration. Different concentrations of P. herba aqueous extract significantly reduced the survival rate, colony formation, number of migrating cells, number of invading cells, and MMP-2 protein expression of MHCC97-H cells, and obviously increased the cell apoptosis rate, the expression levels of Caspase-3, E-cadherin protein and HTR2A-AS1 (P<0.05), which were all concentration-dependent. Overexpression of HTR2A-AS1 evidently decreased the survival rate, colony formation, number of migrating cells, number of invading cells, and MMP-2 protein levels in MHCC97-H cells, while remarkably enhanced the apoptosis rate of cells, P21, Caspase-3, and E-cadherin protein levels and HTR2A-AS1 expression level (P<0.05). Inhibition of HTR2A-AS1 greatly improved the survival rate, the number of clone formation, the number of migrating cells, the number of invading cells and the expression of MMP-2 protein of MHCC97-H cells treated with P. herba aqueous extract, dramatically reducing the cell apoptosis rate, P21, Caspase-3, E-cadherin protein levels and HTR2A-AS1 expression levels (P<0.05). P. herba aqueous extract may inhibit the proliferation, invasion and migration of hepatocellular carcinoma cells by up-regulating the expression of HTR2A-AS1 in hepatocellular carcinoma cells and induce apoptosis.
本研究旨在探讨白花蛇舌草水提物对肝癌细胞增殖、凋亡、侵袭和迁移的作用及机制。用 2.5、5、10mg/mL 白花蛇舌草水提物处理肝癌细胞 MHCC97-H,分别用细胞计数试剂盒(CCK-8)、流式细胞术、平板克隆实验和 Transwell 检测细胞存活率、凋亡、集落形成、侵袭和迁移。实时定量 PCR(qPCR)和 Western blot 分别检测长链非编码 RNA(lncRNA)HTR2A-AS1 和 P21、Caspase-3、E-钙黏蛋白和基质金属蛋白酶-2(MMP-2)蛋白的表达。转染 MHCC97-H 细胞 pcDNA3.1-HTR2A-AS1,或转染 si-HTR2A-AS1 并加用 10mg/mL 白花蛇舌草水提物,评估其对细胞增殖、凋亡、侵袭和迁移的作用。不同浓度白花蛇舌草水提物显著降低 MHCC97-H 细胞存活率、集落形成、迁移细胞数、侵袭细胞数和 MMP-2 蛋白表达,明显增加细胞凋亡率、Caspase-3、E-钙黏蛋白蛋白表达和 HTR2A-AS1 水平(P<0.05),均呈浓度依赖性。过表达 HTR2A-AS1 明显降低 MHCC97-H 细胞存活率、集落形成、迁移细胞数、侵袭细胞数和 MMP-2 蛋白水平,同时显著增强细胞凋亡率、P21、Caspase-3 和 E-钙黏蛋白蛋白水平及 HTR2A-AS1 表达水平(P<0.05)。抑制 HTR2A-AS1 极大地提高了白花蛇舌草水提物处理的 MHCC97-H 细胞的存活率、克隆形成数、迁移细胞数、侵袭细胞数和 MMP-2 蛋白表达,显著降低细胞凋亡率、P21、Caspase-3、E-钙黏蛋白蛋白水平及 HTR2A-AS1 表达水平(P<0.05)。白花蛇舌草水提物可能通过上调肝癌细胞中 HTR2A-AS1 的表达抑制肝癌细胞的增殖、侵袭和迁移,并诱导凋亡。