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Archive > Volume 17 Issue 2 > 2025,17(2):322-339. DOI:https://doi.org/10.1016/j.chmed.2025.02.005 Prev Next

薏苡仁種子提取物聯(lián)合索拉非尼對(duì)HCT116和HepG2細(xì)胞凋亡的促進(jìn)作用

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Enhancement of apoptosis in HCT116 and HepG2 cells by Coix lacryma-jobi var. lacryma-jobi seed extract in combination with sorafenib

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    摘要:
    目的：薏苡是一種備受推崇的亞洲中藥，具有促進(jìn)全身健康和治療多種疾病的雙重功效。雖然單獨(dú)使用時(shí)表現(xiàn)出中等的抗癌效果，但本研究研究了生的和熟的薏苡仁提取物與索拉非尼聯(lián)合使用對(duì)HCT116和HepG2癌細(xì)胞的抗癌潛力。索拉非尼與其他抗癌藥物（包括天然提取物）的聯(lián)合使用，作為開(kāi)發(fā)更有效的癌癥治療的有前途的策略，已經(jīng)引起了極大的關(guān)注。方法：從泰國(guó)當(dāng)?shù)厣虡I(yè)來(lái)源獲得的生(R)和熟(C) CL種子的干粉末，用乙醇(E)、二氯甲烷(D)、乙酸乙酯(A)和水(W)提取和分離，得到8個(gè)餾分：CLRE、CLCE、CLRD、CLCD、CLRA、CLCA、CLRW和CLCW。對(duì)生、熟種子提取物中薏苡醇的含量進(jìn)行量化，并以每克提取物中薏苡醇的含量表示。使用MTT法評(píng)估這些組分對(duì)HCT116和HepG2細(xì)胞的細(xì)胞毒性作用。顯示最顯著的細(xì)胞毒性反應(yīng)的組分與索拉非尼聯(lián)合評(píng)估其協(xié)同作用。評(píng)估細(xì)胞凋亡誘導(dǎo)和線粒體膜電位（MMP），并通過(guò)分析活性氧（ROS）的產(chǎn)生和抗氧化蛋白的表達(dá)水平探討細(xì)胞凋亡的潛在機(jī)制。此外，我們還研究了聯(lián)合治療對(duì)磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B (AKT)/雷帕霉素（mTOR）通路的機(jī)制靶點(diǎn)的影響。結(jié)果：每克CLCE和CLCD提取物的薏苡醇含量分別為7.02和9.69 lg，高于CLRE和CLRD（分別為2.66和5.96 lg）。在本研究條件下，CLRA、CLRW和CLCW餾分中的薏苡醇含量均未檢測(cè)到。與HCT116和HepG2細(xì)胞孵卵24小時(shí)和48小時(shí)后，所有提取物的IC50值均超過(guò)1 mg/mL，表明單獨(dú)使用時(shí)細(xì)胞毒性有限。選擇CLRD和CLCD組分進(jìn)行聯(lián)合研究，濃度為1mg/mL，與低于IC50濃度的索拉非尼聯(lián)合使用，以盡量減少其副作用。該組合通過(guò)提高ROS水平和降低超氧化物歧化酶2和過(guò)氧化氫酶的表達(dá)，顯著增加細(xì)胞毒性，誘導(dǎo)HCT116和HepG2細(xì)胞凋亡。此外，聯(lián)合治療下調(diào)了PI3K/AKT/mTOR通路，表明其具有靶向抗癌機(jī)制。結(jié)論：CLCD聯(lián)合索拉非尼作為未來(lái)抗癌治療策略具有顯著的潛力。這種CL種子提取物，在泰國(guó)種植和商業(yè)化，顯示出希望作為一種天然補(bǔ)充劑，以提高化療的療效，在即將到來(lái)的臨床抗癌應(yīng)用。

    Abstract:
    Objective: Coix lacryma-jobi, a highly regarded Asian herb widely used in traditional Chinese medicine, is recognized for its dual benefits in promoting overall health and treating various diseases. While it exhibits moderate anticancer efficacy when used alone, this study investigated the enhanced anticancer potential of raw and cooked Coix lacryma-jobi var. lacryma-jobi (CL) seed extracts in combination with sorafenib against HCT116 and HepG2 cancer cell lines. The combination of sorafenib with other anticancer agents, including natural extracts, has garnered significant attention as a promising strategy for developing more effective cancer therapies. Methods: Dry powders of raw (R) and cooked (C) CL seeds, obtained from a local commercial source in Thailand, were extracted and fractionated using ethanol (E), dichloromethane (D), ethyl acetate (A), and water (W) to produce eight fractions: CLRE, CLCE, CLRD, CLCD, CLRA, CLCA, CLRW, and CLCW. The coixol content in raw and cooked seed extracts was quantified and expressed as lg of coixol per gram of extract. The cytotoxic effects of these fractions were evaluated against HCT116 and HepG2 cells using the MTT assay. Fractions demonstrating the most significant cytotoxic responses were combined with sorafenib to evaluate their synergistic effects. Apoptosis induction and mitochondrial membrane potential (MMP) were assessed, and the underlying mechanism of apoptosis was explored by analyzing reactive oxygen species (ROS) generation and antioxidant protein expression levels. Additionally, the combination treatment’s effect on the phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) pathway was investigated. Results: One gram of CLCE and CLCD extracts contained higher coixol levels (7.02 lg and 9.69 lg, respectively) compared to CLRE and CLRD (2.66 lg and 5.96 lg, respectively). Coixol content in CLRA, CLRW, and CLCW fractions was undetectable under the study conditions. All extract fractions exhibited IC50 values exceeding 1 mg/mL after 24- and 48-hour incubations with HCT116 and HepG2 cells, indicating limited cytotoxicity when used independently. CLRD and CLCD fractions were selected for combination studies at a concentration of 1 mg/mL, combined with sub-IC50 concentrations of sorafenib to minimize its side effects. This combination significantly increased cytotoxicity, inducing apoptosis in HCT116 and HepG2 cells by elevating ROS levels and reducing the expression of superoxide dismutase 2 and catalase. Furthermore, the combination treatment downregulated the PI3K/AKT/mTOR pathway, indicating a targeted anticancer mechanism. Conclusion: The combination of CLCD with sorafenib demonstrates significant potential as a strategy for future anticancer therapies. This CL seed extract, cultivated and commercially available in Thailand, shows promise as a natural supplement to enhance the efficacy of chemotherapy in upcoming clinical anticancer applications.

    關(guān)鍵詞:
    細(xì)胞凋亡;薏苡仁;HCT116;HepG2;PI3K/AKT/mTOR通路;活性氧;種子提取物 ;索拉非尼

    Keywords:


    Project Supported:
    This work was supported by the National Science Research and Innovation Fund (NSRF) of Thailand (No. R2565B045), the Global and Frontier Research University Fund, (No. R2567C003), Naresuan University, Thailand. The funders played no part in study design, data collection and analysis, publication decisions, or manuscript writing. This manuscript has been proofread and edited by Mr. Olalekan Israel Aiikulola, Lecturer (Special Knowledge and Abilities), Faculty of Medical Science, Naresuan University, Thailand.

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