补骨脂乙素通过调控Sirtuins相关信号通路抑制非小细胞肺癌进程

目的：观察补骨脂乙素对人非小细胞肺癌增殖、凋亡和周期的影响,探究补骨脂乙素抑制H460细胞活性的分子机制。方法：采用MTT法检测补骨脂乙素对H460细胞增殖活力的影响;流式细胞术检测补骨脂乙素对H460细胞周期阻滞的影响;Annexin V-FITC/PI染色法探究补骨素乙素对H460细胞凋亡的影响;免疫印迹法检测细胞内信号通路中关键蛋白的表达情况。结果：与空白对照组相比,补骨脂乙素(60μM)和SirReal2(50μM)给药组明显抑制细胞增殖,促使细胞周期阻滞在G0/G1期,诱导细胞内ROS生成以及细胞凋亡,并且明显抑制了SIRT1和SIRT2的去乙酰化活性,促使其相关蛋白C-myc,Cyclin D1,Bcl-2表达减少,Bax和乙酰化的α-tubulin表达明显增多。结论：补骨脂乙素对H460细胞的增殖和细胞周期具有明显抑制作用,并且还会诱导ROS生成从而导致细胞凋亡,其分子机制可能是通过影响细胞内Sirtuins相关的信号通路实现的。     

人参皂苷compound K对慢性粒细胞白血病K562细胞增殖抑制和凋亡诱导作用

目的探讨人参皂苷compound K（CK）对慢性粒细胞白血病K562细胞增殖抑制和凋亡的影响。方法应用MTT法检测CK对K562细胞增殖的影响,用流式细胞仪分析细胞周期,DNA Ladder实验观察CK对K562细胞凋亡的影响。结果人参皂苷CK能显著抑制K562细胞的增殖,呈浓度依赖性,细胞阻滞于G2/M期;CK能诱导K562细胞的凋亡,呈浓度依赖性。结论人参皂苷CK具有抑制K562细胞增殖和诱导凋亡作用。     

全反式维甲酸对小细胞肺癌细胞的分化诱导作用研究

目的探讨全反式维甲酸(all-trans-retinoic acid,ATRA)对小细胞肺癌细胞的分化诱导作用。方法选择NCI-H446细胞株MTT法检测细胞体外增殖能力,电镜下观察形态,流式细胞仪进行细胞周期分析。RT-PCR法分析维甲酸受体亚型基因。结果全反式维甲酸诱导小细胞肺癌NCI-H446细胞株,细胞增殖能力下降,凋亡增加,更多的细胞被阻止于G1/G0期。结论全反式维甲酸诱导分化能有效抑制小细胞肺癌NCI-H446细胞株的增殖,促进凋亡。     

丹参酮衍生物对K562细胞的生长抑制及诱导凋亡作用研究

目的研究丹参酮衍生物对人红白血病细胞株K562的生长抑制以及诱导凋亡作用。方法MTT比色法检测不同浓度丹参酮1、丹参酮2和丹参酮B对K562细胞的增殖抑制作用;PI单染法检测3者对K562细胞周期的影响;An-nexin V/PI双染法检测3者对K562细胞诱导凋亡的作用。结果丹参酮1和丹参酮B能够抑制K562细胞增殖,IC50分别为5.22和15.11μmol·L-1,且分别在2.5~10μmol·L-1和10~40μmol·L-1剂量范围内,G0/G1期细胞比例的增加及早期凋亡细胞百分率的提高均呈剂量相关性;丹参酮2在100μmol·L-1的剂量下,对K562细胞的抑制率仅为27.8%,无诱导其凋亡作用,但可以使G0/G1期细胞增多。结论丹参酮1和丹参酮B抑制K562细胞增殖,阻滞其于G0/G1期并诱导细胞凋亡;丹参酮2对K562细胞没有明显生长抑制及诱导凋亡作用,但可将其阻滞于G0/G1期。     

葫芦素B通过激活线粒体途径诱导人肺癌NCI-H460细胞凋亡

目的探讨葫芦素B对人肺癌NCI-H460细胞增殖及凋亡的影响,并探讨其机制。方法体外培养NCI-H460细胞,MTT法观察葫芦素B对细胞增殖的影响,倒置相差显微镜观察细胞形态的变化,采用流式细胞术检测细胞凋亡率。采用罗丹明123(Rhodamine 123)染色,通过流式细胞仪检测线粒体膜电位(△ψm),采用Western blot方法检测线粒体内和胞浆内的细胞色素C。结果葫芦素B对NCI-H460细胞的增殖有抑制作用,且呈剂量、时间依赖性;葫芦素B处理组细胞形态发生显著改变,细胞皱缩、变圆,可见胞核染色质浓缩;流式细胞仪检测结果显示葫芦素B诱导NCI-H460细胞凋亡,呈剂量依赖性。葫芦素B处理后线粒体膜电位显著下降,细胞色素C由线粒体释放到胞浆中。结论葫芦素B可以抑制人肺癌NCI-H460细胞的增殖并诱导细胞凋亡,线粒体途径参与葫芦素B诱导的NCI-H460细胞凋亡。     

PD98059对HL60细胞Ras-MEK1/2-ERK1/2信号转导影响的研究

目的探讨MEK1抑制剂PD98059对HL60细胞Ras-MEK1/2-ERK1/2信号转导以及细胞增殖的影响。方法四唑盐比色试验(MTT)法检测PD98059对HL60细胞增殖的抑制作用,流式细胞术观察PD98059对HL60细胞凋亡和G0/G1期阻滞的影响,半定量反转录-聚合酶链反应(RT-PCR)测定PD98059对HL60细胞ERK2、p27、Skp2基因表达的影响,免疫组织化学SP法检测ERK2、p27、Skp2蛋白表达的改变情况。结果 PD98059能够抑制HL60细胞的生长,该抑制作用具有浓度及时间依赖性(P<0.05)。PD98059能够促进HL60细胞凋亡,该作用具有浓度及时间依赖性(P<0.05);PD98059作用HL60细胞48h,随着浓度的增加G0/G1期阻滞增强(P<0.05)。PD98059可使HL60细胞ERK2、Skp2的mRNA及蛋白表达量减少,p27的mR-NA及蛋白表达量增加(P<0.05)。结论 PD98059能够抑制HL60细胞的生长,诱导细胞发生G0/G1期阻滞,促进其凋亡,这可能是通过PD98059阻断Ras-MEK1/2-ERK1/2信号转导,影响了ERK2、Skp2、p27等的表达而实现的。     

美洲大蠊提取物对两株人肺癌细胞的影响

目的:探讨美洲大蠊提取物(CⅡ-3)对两株人肺癌细胞(NCI-H446、NCI-H460)的细胞毒活性及细胞周期的影响。方法:采用改良MTT法检测了CⅡ-3对NCI-H446和NCI-H460人肺癌细胞的细胞毒活性,流式细胞术检测CⅡ-3对2株细胞凋亡、坏死率及细胞周期的的影响。结果:CⅡ-3对NCI-H446和NCI-H460细胞的IC50分别为(48.98±6.63)μg.mL-1、(33.00±10.57)μg.mL-1。对NCI-H446和NCI-H460细胞凋亡、坏死率的影响具有一定的剂量依赖性。对NCI-H446细胞,25,50μg.mL-1CⅡ-3诱导细胞的不同时期发生阻滞,75μg.mL-1CⅡ-3主要诱导细胞发生坏死;对NCI-H460细胞,20,80μg.mL-1CⅡ-3主要诱导细胞S期、G2/M期发生阻滞,40μg.mL-1CⅡ-3主要诱导细胞S期发生阻滞。结论:美洲大蠊提取物CⅡ-3对2株人肺癌细胞有一定的细胞毒活性,且对2株细胞的凋亡、坏死率和细胞周期分布有一定的影响。     

分解前后白术挥发油对细胞凋亡及细胞周期影响的研究

目的研究分解前、后白术挥发油对人类卵巢癌细胞SKOV-3细胞形态、生长抑制率、细胞凋亡及细胞周期的影响。方法使用倒置显微镜及采用四甲基偶氮唑蓝(MTT)法和流式细胞术,观察不同浓度分解前、后白术挥发油作用SKOV-3细胞不同时间后,对细胞形态、生长抑制率、细胞凋亡及细胞周期的影响。结果镜下可见凋亡小体。分解前、后白术挥发油各浓度样品作用细胞时间72h,或100,200mg/L作用细胞24,48h时,对细胞增殖均有显著影响(P<0.01)。浓度12.5,25,50mg/L分解后白术挥发油作用细胞24,48h时,对细胞增殖有显著影响(P<0.05)。浓度25,50mg/L分解前白术挥发油作用细胞24,48,72h时的细胞增殖抑制率明显高于分解后挥发油的抑制率,差异有统计学意义。随着分解前、后挥发油作用浓度的增加,分解前白术挥发油早期细胞凋亡率增高,晚期凋亡率减少;分解后白术挥发油早期细胞凋亡率、晚期细胞凋亡率增加;分解前挥发油G0/G1期和S期的细胞比例降低,G2/M期的细胞比例升高;分解后挥发油G0/G1期和G2/M期的细胞比例降低,S期的升高。结论分解前、后白术挥发油对SKOV-3细胞均具有杀伤抑制作用,其作用与时间和剂量呈正相关。在一定浓度范围,分解前白术挥发油对SKOV-3细胞增殖影响明显大于分解后。分解前、后白术挥发油均具有诱导细胞凋亡作用。分解前白术挥发油主要在凋亡早期发挥作用,分解后的挥发油在凋亡早期、晚期均发挥作用;分解前挥发油可将细胞阻滞在G2/M期,分解后可将细胞阻滞在S期。     

血管紧张素Ⅱ受体抑制剂对人卵巢癌细胞抑制作用的研究

目的探讨血管紧张素Ⅱ1型受体拮抗剂(氯沙坦)对人卵巢癌HO89101细胞的体外生长的影响。方法采用体外实验的方法将不同浓度的Losartan作用于HO8910细胞后,运用四甲基偶氮唑蓝(MTT)法观察氯沙坦对HO8910细胞增殖的抑制作用,并采用流式细胞仪分析氯沙坦对HO8910细胞增殖周期的影响。结果①不同浓度的氯沙坦对HO8910细胞有抑制其生长的作用(P<0.05),并呈现剂量依赖性关系;②细胞周期分析显示氯沙坦使HO8910 G0/G1期细胞明显增加,而S和G2/M期的细胞减少,说明细胞被阻滞在G0/G1期,细胞增殖被抑制。结论氯沙坦可以抑制HO8910细胞的体外增殖,细胞被阻滞在G0/G1期,诱导细胞进行程序性死亡。     

棘豆素对人雄激素非依赖性前列腺癌 PC-3细胞的作用研究

【】目的: 研究棘豆素(2′, 4′-dihydroxychalcone,TFC)对人雄激素非依赖性前列腺癌PC-3细胞的作用,并探讨其作用机制。方法：CCK-8法检测药物对细胞增殖的影响,Hoechst 33258染色、荧光显微镜观察细胞的形态学变化,流式细胞术分析细胞凋亡率和细胞周期的变化, Western blot检测药物对肿瘤细胞中P27kip1和PTEN蛋白表达的影响。结果：TFC能显著抑制PC-3细胞的增殖,细胞呈现凋亡形态学改变,细胞凋亡率明显增加,细胞周期阻滞于G0/G1期。此外,P27kip1和PTEN蛋白表达量明显增加。结论：TFC通过诱导PC-3细胞凋亡抑制肿瘤细胞增殖,可能与其G0/G1细胞周期阻滞,以及PTEN 和P27Kip1蛋白表达上调有关。     

冬虫夏草菌丝体水提醇沉物体外抗肿瘤活性研究

目的 研究冬虫夏草菌丝体水提醇沉物的体外抗肿瘤活性及其机制。方法 采用热水浸提-醇沉法得到水提物,高效液相色谱仪测定水提醇沉物中腺苷的量;采用MTT法测定其抗肿瘤活性,利用流式细胞仪结合碘化丙锭染色法检测其对细胞周期的抑制。结果 实验表明水提醇沉物能抑制人肝癌HepG2细胞株及大细胞肺癌NCI-H460细胞株的增殖,并呈浓度相关性;半数抑制浓度（IC₅₀）值分别为（1.49±0.19）和（1.67±0.27）mg/mL。细胞周期分析表明,水提醇沉物分别阻滞HepG2及NCI-H460细胞周期于G₂/M期、S期,并可诱导上述两种细胞发生凋亡。结论 冬虫夏草水提醇沉物通过阻滞HepG2及NCI-H460细胞周期循环,诱导其凋亡,从而表现出良好的增殖抑制活性。为深入研究冬虫夏草菌丝体水提醇沉物抗肿瘤的机制提供了实验证据。     

Carnosic acid exhibits antiproliferative and proapoptotic effects in tumoral NCI-H460 and nontumoral IMR-90 lung cells

ABSTRACT

Carnosic acid (CA) is a phenolic diterpene with many important biological activities including antimicrobial, antioxidant, anti-inflammatory properties, and anti-proliferative properties. The aim of the present study was to investigate cytotoxic activity, cell cycle, apoptotic, and molecular effects attributed to CA in non-tumoral IMR-90 (human fetal lung fibroblasts), as well as tumoral NCI-H460 (human non–small-cell lung cancer) cell lines. Cell proliferation was evaluated by Real-Time Cell Analysis system, while apoptosis and cell cycle were assessed using flow cytometry. RT-qPCR was used to estimate the relative expression of genes involved in cell cycle regulation, DNA damage and repair, and apoptosis induction. CA inhibited proliferation of IMR-90 and NCI-H460 cells via cell cycle arrest at G₀/G₁ and G₂/M phases, according to the treatment concentration. The mRNA levels of genes encoding cyclins A2, B1, and B2 were downregulated in response to CA treatment of IMR-90 cells. Apoptosis was induced and proapoptotic gene PUMA was upregulated in both cell lines. mRNA levels of genes ATR, CCND1, CHK1, CHK2, MYC, GADD45A, H2AFX, MTOR, TP53, and BCL2, CASP3 were not markedly changed following CA treatments. Although CA exerted antiproliferative activity against NCI-H460 tumor cells, this phytochemical induced toxic effects in non-tumoral cells, and thus needs to be considered carefully prior to pharmacological use therapeutically.     

人参皂苷Rg3对乳腺癌MCF-7细胞增殖和侵袭的影响

目的 观察人参皂苷Rg3对雌激素受体阳性的乳腺癌细胞MCF-7增殖和侵袭的影响,并探讨其可能的作用机制。方法 采用MTT法检测细胞的增殖能力,流式细胞仪分析细胞周期分布以及凋亡比率,通过Transwell小室观察细胞侵袭力,RT-PCR法检测细胞中的MMP-9 mRNA的表达。结果 与对照组相比,人参皂苷Rg3能显著抑制MCF-7细胞的增殖;G₀/G₁期及S期细胞比例减少,而G₂/M期细胞比例显著增加;同时细胞凋亡比率亦明显提升,而细胞侵袭指数降低,且呈现良好的剂量、时间依赖性。同时人参皂苷Rg3还能显著抑制细胞中MMP-9 mRNA的表达水平(P<0.05)。结论 人参皂苷Rg3能抑制MCF-7细胞的增殖和侵袭,其作用机制可能与其能降低MMP-9基因的表达有关。     

杠柳毒苷体外抑制肝癌细胞和乳腺癌细胞增殖的实验研究

目的 探讨杠柳毒苷在体外对人乳腺癌MDA-MB-468细胞和人肝癌HepG2细胞增殖的影响。方法 MTT法观察杠柳毒苷对人乳腺癌MDA-MB-468细胞和人肝癌HepG2细胞增殖的抑制作用,流式细胞术观察杠柳毒苷对两种肿瘤细胞的细胞增殖周期作用。结果 与对照组比较,杠柳毒苷能明显抑制两种肿瘤细胞的增殖,其抑制率与药物浓度和作用时间呈正相关。流式细胞仪检测发现,杠柳毒苷对乳腺癌MDA-MB-468细胞和肝癌HepG2细胞持续作用24 h后,可以使G₀/G₁期细胞增多,G₂/M期细胞减少。结论 杠柳毒苷具有抑制乳腺癌MDA-MB-468细胞和肝癌HepG2细胞增殖的作用,并可将乳腺癌MDA-MB-468细胞和肝癌HepG2细胞的细胞生长周期阻滞在G₀/G₁期。     

冬虫夏草菌丝体水提醇沉物体外抗肿瘤活性研究

目的研究冬虫夏草菌丝体水提醇沉物的体外抗肿瘤活性及其机制。方法采用热水浸提一醇沉法得到水提物,高效液相色谱仪测定水提醇沉物中腺苷的量;采用MTT法测定其抗肿瘤活性,利用流式细胞仪结合碘化丙锭染色法检测其对细胞周期的抑制。结果实验表明水提醇沉物能抑制人肝癌HepG2细胞株及大细胞肺癌NCI．H460细胞株的增殖,并呈浓度相关性;半数抑制浓度（IC50）值分别为（1．49±0．19）和（1．67±0．27）mg／mL。细胞周期分析表明,水提醇沉物分别阻滞HepG2及NCI．H460细胞周期于G,／M期、S期,并可诱导上述两种细胞发生凋亡。结论冬虫夏草水提醇沉物通过阻滞HepG2及NCI-H460细胞周期循环,诱导其凋亡,从而表现出良好的增殖抑制活性。为深入研究冬虫夏草菌丝体水提醇沉物抗肿瘤的机制提供了实验证据。     

Flavonoids from Vitex trifolia L. inhibit cell cycle progression at G2/M phase and induce apoptosis in mammalian cancer cells

Six flavonoids, persicogenin (1), artemetin (2), luteolin (3), penduletin (4), vitexicarpin (5) and chrysosplenol-D (6), have been isolated for the first time as new cell cycle inhibitors from Vitex trifolia L., a Chinese folk medicine used to treat cancers, through a bioassay-guided separation procedure. They were identified by spectroscopic methods. The inhibitory effects of 1–6 on the proliferation of mammalian cancer cells have been evaluated by the SRB (sulforhodamine B) method and their effects on cell cycle and apoptosis investigated by flow cytometry with the morphological observation under light microscope and by agarose-gel electrophoresis to detect internucleosomal DNA fragmentation. Compounds 1–6 inhibited the proliferation of mouse tsFT210 cancer cells with the IC₅₀s (μg?ml^?1) > 100 (inhibition rate at 100?μg?ml^?1, 47.9%) for 1, >100 (inhibition rate at 100?μg?ml^?1, 49.6 %) for 2, 10.7 for 3, 19.8 for 4, 0.3 for 5, and 3.5 for 6. Flow cytometric investigations for 1–6 demonstrated that 1–5 mainly inhibited cell cycle at the G₂/M phase in a dose-dependent manner with a weak induction of apoptosis on the tsFT210 cells, while 6 induced mainly apoptosis of the same tsFT210 cells also in a dose-dependent manner together with a weak inhibition of the cell cycle at the G₀/G₁ and G₂/M phases, demonstrating that 1–6 exert their anti-proliferative effect on tsFT210 cells through inhibiting cell cycle and inducing apoptosis. In contrast to the cell cycle G₂/M phase inhibitory main effect on tsFT210 cells, 5 induced mainly apoptosis on human myeloid leukemia K562 cells with a weak inhibition of the cell cycle at the G₂/M phase. The present result provides flavonoids 1–6 as new cell cycle inhibitors and 1 and 4 as new anticancer flavonoids, which not only provide the first example of cell cycle G₂/M phase inhibitory and apoptosis-inducing constituents of V. trifolia L. but also explain the use of Vitex trifolia L. by Chinese people to treat cancers.     

Celecoxib in combination with retinoid CD437 inhibits melanoma A375 cell in vitro

This study aimed to investigate the effects of celecoxib, synthetic retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalenecarboxylicacid (CD437) and the combination of the two on cell proliferation, apoptosis, and cycle arrest of human malignant melanoma A375 cells. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay (MTT assay) was applied to determine the anti-proliferative effects of the drugs on human malignant melanoma A375 cells. Flow cytometry was performed to investigate the influence of the drugs on cell cycle and cell apoptosis. Both celecoxib and CD437 could inhibit the growth of human malignant melanoma A375 cells in a dose-dependent manner. Celecoxib at 80 μmol/L inhibited proliferation, induced apoptosis and G₂/M cell cycle arrest of human malignant melanoma A375 cells after treatment for 24 h [proliferation inhibiting rate: (50.2 ± 2.51)%, apoptosis rate: (35.91 ± 1.80)%]. CD437 at 10 μmol/L inhibited proliferation, induced apoptosis and G₀/G₁ cell cycle arrest of human malignant melanoma A375 cells after treatment for 24 h [proliferation inhibiting rate: (58.6 ± 2.38)%, apoptosis rate: (28.03 ± 0.77)%]. Celecoxib in combination with CD437 could significantly enhance the effects of inhibiting proliferation and inducing apoptosis of human malignant melanoma A375 cells 24 h after treatment compared with the drug alone [proliferation inhibiting rate: (68.92 ± 1.72)%, apoptosis rate: (42.09 ± 1.05)%, both P<0.05] and decrease the proportion of the S phase in the cell cycle. Celecoxib could inhibit the growth of human malignant melanoma A375 cells by inducing apoptosis and G₂/M cycle arrest. CD437 could inhibit the growth of human malignant melanoma A375 cells by inducing apoptosis and G₀/G₁ cycle arrest. Celecoxib exhibited additive effects with CD437 on retarding the growth and inducing apoptosis of human malignant melanoma A375 cells. Celecoxib in combination with CD437 may become an effective method for prevention and treatment of human melanoma.     

Anti-proliferative properties of commercial Pelargonium sidoides tincture,with cell-cycle G0/G1 arrest and apoptosis in Jurkat leukaemia cells

Context Pelargonium sidoides DC (Geraniaceae) is an important medicinal plant indigenous to South Africa and Lesotho. Previous studies have shown that root extracts are rich in polyphenolic compounds with antibacterial, antiviral and immunomodulatory activities. Little is known regarding the anticancer properties of Pelargonium sidoides extracts.

Objective This study evaluates the anti-proliferative effects of a Pelargonium sidoides radix mother tincture (PST).

Materials and methods The PST was characterized by LC-MS/MS. Anti-proliferative activity was evaluated in the pre-screen panel of the National Cancer Institute (NCI-H460, MCF-7 and SF-268) and the Jurkat leukaemia cell line at concentrations of 0–150?μg/mL. The effect on cell growth was determined with sulphorhodamine B and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays after 72?h. The effect on cell cycle and apoptosis induction in Jurkat cells was determined by flow cytometry with propidium iodide and Annexin V: fluorescein isothiocyanate staining.

Results Dihydroxycoumarin sulphates, gallic acid as well as gallocatechin dimers and trimers were characterized in PST by mass spectrometry. Moderate anti-proliferative effects with GI₅₀ values between 40 and 80?μg/mL were observed in the NCI-pre-screen panel. Strong activity observed with Jurkat cells with a GI₅₀ value of 6.2?μg/mL, significantly better than positive control 5-fluorouracil (GI₅₀ value of 9.7?μg/mL). The PST arrested Jurkat cells at the G₀/G₁ phase of the cell cycle and increased the apoptotic cells from 9% to 21%, while the dead cells increased from 4% to 17%.

Conclusion We present evidence that P. sidoides has cancer cell type-specific anti-proliferative effects and may be a source of novel anticancer molecules.     

洛伐他汀联合顺铂对人肝癌HepG2细胞生物学特性的影响及其机制

目的 研究洛伐他汀单用或与化疗药物顺铂联用时对人肝癌HepG2细胞生物学特性的影响,初步探索其抗肿瘤作用机制。方法 不同浓度洛伐他汀、洛伐他汀联合顺铂处理细胞 48 h后,CCK-8法检测HepG2细胞的增殖抑制作用,金氏公式计算联合应用效果;平板克隆形成实验评价药物作用于肝癌细胞的远期效应;划痕实验检测药物对细胞迁移能力的影响;Transwell小室法检测药物对细胞侵袭能力的影响;流式细胞术检测药物处理后细胞周期和凋亡情况;蛋白印迹技术（Western-blot）检测凋亡相关蛋白Bcl-2、Bax、Caspase-3的表达水平变化。结果 洛伐他汀呈浓度依赖性抑制HepG2细胞的增殖,金氏公式计算结果显示洛伐他汀可协同增强顺铂的抗肿瘤作用;平板克隆形成实验结果表明洛伐他汀能显著抑制HepG2细胞克隆形成率;划痕实验提示洛伐他汀能显著降低肝癌细胞的迁移率;Transwell小室侵袭实验结果发现洛伐他汀能明显减少穿膜细胞数量;流式细胞检测发现洛伐他汀可引起G0/G1期细胞增加,S期细胞减少,细胞凋亡率增加;Western-blot检测结果显示洛伐他汀可下调Bcl-2蛋白表达,同时上调Bax和Caspase-3蛋白表达。结论 洛伐他汀可明显抑制HepG2细胞增殖、迁移和侵袭能力,与顺铂联用可增强顺铂体外抗肿瘤效果,通过线粒体途径诱导肿瘤细胞凋亡是其可能的抗肿瘤作用机制之一。     

Chan-Yu-Bao-Yuan-Tang induces apoptosis in NSCLC and SCLC cell lines via a mitochondria-mediated pathway

1. Previous clinical studies have shown efficacy and safety of the traditional Chinese medicinal herb extract Chan-Yu-Bao-Yuan-Tang (CYBYT) in lung cancer patients.

2. The effects of CYBYT on proliferation and apoptosis in the non-small cell lung cancer cell line NCI-H460 and the small cell lung cancer cell line NCI-H446 were investigated in vitro.

3. CYBYT significantly induced antiproliferative effects of NCI-H460 and NCI-H446 cells in a concentration- and time-dependent manner. The IC₅₀ values in NCI-H460 cells were 94.37 µg/mL (24?h) and 20.89 µg/mL (48?h), whereas in NCI-H446 cells IC₅₀ values were 214.72 µg/mL (24?h) and 114.58 µg/mL (48?h). Annexin V–FITC/PI staining showed CYBYT could significantly induce apoptosis in NCI-H460 and NCI-H446 cells, and the total apoptosis rates were positively correlated with the concentration and time of CYBYT treatment. Furthermore, treatment with the broad-spectrum caspase inhibitor (z-VAD-fmk) effectively protected NCI-H460 and NCI-H446 cells against CYBYT-triggered apoptosis. The apoptotic processes involved were a marked decrease in antiapoptotic protein Bcl-2 and an increase in proapoptotic protein Bax. The release of mitochondrial cytochrome c into the cytosol was also observed, which, in turn, resulted in the activation of caspase-9 and caspase-3.

4. CYBYT exerts antiproliferative and growth inhibition effects on NCI-H460 and NCI-H446 cells through the mitochondrial caspase-dependent cell death pathway.