Sitthisuk Pornnapa, Poorahong Watcharaporn, Innajak Sukanda, Krajarng Aungkana, Samosorn Siritron, Watanapokasin Ramida
Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand.
Department of Biochemistry, Faculty of Medicine, Bangkok Thonburi University, Bangkok 10170, Thailand.
Pharmaceuticals (Basel). 2025 Mar 21;18(4):441. doi: 10.3390/ph18040441.
(MS) is a Thai herb used in traditional medicine. Previous studies have reported the antiproliferative effects of its constituents in various cancer cell lines. However, the effects of MS extract on cytotoxicity and molecular mechanisms of apoptosis induction in HCT116 colon cancer cells have not been fully explored. The cytotoxic effect of MS extract on HCT116 cells was assessed using the MTT assay. MS extract increased cytotoxicity in a concentration-dependent manner. It also induced nuclear morphological changes and disrupted the mitochondrial membrane potential (m), as assessed by Hoechst 33342 and JC-1 staining, respectively. These findings indicated that MS extract induced apoptosis, which was further confirmed by flow cytometry showing an increase in the sub-G1 phase. To investigate the expression of signaling proteins, Western blot analysis was conducted. The results showed that MS extract activated caspase activity (caspase-8, -9, and -7) and inhibited PARP activity. Additionally, MS extract upregulated pro-apoptotic proteins (tBid, Bak, and cytochrome c) while downregulating anti-apoptotic proteins (Bcl-2 and Bcl-xL). Mechanistic studies revealed that MS extract activated MAPK pathways while inactivating the PI3K/Akt/NF-κB and GSK-3β/β-catenin pathways. Notably, MS extract also inhibited V-ATPases, as evaluated by acridine orange staining and Western blot analysis. Our findings suggest that MS extract induces apoptosis via the activation of both intrinsic and extrinsic pathways associated with the key signaling pathways. Therefore, MS extract shows potential as a therapeutic agent for colon cancer.
MS是一种用于传统医学的泰国草药。先前的研究报道了其成分在各种癌细胞系中的抗增殖作用。然而，MS提取物对HCT116结肠癌细胞的细胞毒性作用及诱导细胞凋亡的分子机制尚未得到充分研究。使用MTT法评估MS提取物对HCT116细胞的细胞毒性作用。MS提取物以浓度依赖性方式增加细胞毒性。通过Hoechst 33342和JC-1染色评估，它还诱导了核形态变化并破坏了线粒体膜电位（m）。这些发现表明MS提取物诱导了细胞凋亡，流式细胞术进一步证实了这一点，显示亚G1期增加。为了研究信号蛋白的表达，进行了蛋白质免疫印迹分析。结果表明，MS提取物激活了半胱天冬酶活性（半胱天冬酶-8、-9和-7）并抑制了PARP活性。此外，MS提取物上调了促凋亡蛋白（tBid、Bak和细胞色素c），同时下调了抗凋亡蛋白（Bcl-2和Bcl-xL）。机制研究表明，MS提取物激活了MAPK通路，同时使PI3K/Akt/NF-κB和GSK-3β/β-连环蛋白通路失活。值得注意的是，通过吖啶橙染色和蛋白质免疫印迹分析评估，MS提取物还抑制了V-ATP酶。我们的研究结果表明，MS提取物通过激活与关键信号通路相关的内在和外在途径诱导细胞凋亡。因此，MS提取物显示出作为结肠癌治疗剂的潜力。
