Gleditsia sinensis thorn extract inhibits human colon cancer cells: the role of ERK1/2, G2/M‐phase cell cycle arrest and p53 expression

The thorns of Gleditsia sinensis are used as a medicinal herb in China and Korea. However, the mechanisms responsible for the antitumor effects of the water extract of Gleditsia sinensis thorns (WEGS) remain unknown. HCT116 cells treated with the WEGS at a dose of 800 μg/mL (IC₅₀) showed a significant decrease in cell growth and an increase in cell cycle arrest during the G2/M‐phase. G2/M‐phase arrest was correlated with increased p53 levels and down‐regulation of the check‐point proteins, cyclinB1, Cdc2 and Cdc25c. In addition, treatment with WEGS induced phosphorylation of extracellular signal‐regulated kinase (ERK), p38 MAP kinase and JNK (c‐Jun N‐terminal kinases). Moreover, inhibition of ERK by treatment of cells with the ERK‐specific inhibitor PD98059 blocked WEGS‐mediated p53 expression. Similarly, blockage of ERK function in the WEGS‐treated cells reversed cell‐growth inhibition and decreased cell cycle proteins. Finally, in vivo WEGS treatment significantly inhibited the growth of HCT116 tumor cell xenografts in nude mice with no negative side effects, including loss of body weight. These results describe the molecular mechanisms whereby the WEGS might inhibit proliferation of colon cancer both in vitro and in vivo, suggesting that WEGS has potential as an anticancer agent for the treatment of malignancies. Copyright © 2010 John Wiley & Sons, Ltd.     

Effect of Orostachys japonicus on cell growth and apoptosis in human hepatic stellate cell line LX2

Orostachys japonicus (O. japonicus), used to treat diseases such as various cancers, gastric ulcers, fever, hepatitis, arthritis, eczema, for hemostasis, and intoxication in folk medicine, has been an important constituent in many herbal formulae. We demonstrated that the water extract of O. japonicus led to growth inhibition of LX2 cells by inducing apoptosis through the caspase activation, related to the MAPK pathway. O. japonicus inhibited proliferation of LX2 cells in a dose- and time-dependent manner, increased the apoptosis fraction at cell cycle progression with an accompanying DNA fragmentation, and resulted in a significant decrease in Bcl-2 and an increase in Bax mRNA levels. Exposure of LX2 cells to O. japonicus induced caspase-3 activation, however when the LX2 cells were also treated with the pan-caspase inhibitor Z-VAD-FMK and the caspase-3 inhibitor Z-DEVE-FMK, apoptosis was blocked. O. japonicus inhibited anti-apoptotic Mcl-1 protein and MEK/ERK phosphorylation in LX2 cells. The results indicate that O. japonicus inhibits the cell growth of LX2 cells by inducing apoptosis through caspase activity. O. japonicus down-regulated Mcl-1 protein levels and inhibited the phosphorylation of MEK/ERK, suggesting that it mediates cell death in LX2 cells through the down-regulation of Mcl-1 protein via a MEK/ERK-independent pathway.     

F1012‐2 inhibits the growth of triple negative breast cancer through induction of cell cycle arrest,apoptosis, and autophagy

Sesquiterpene lactones (SLs) are plant‐derived constituents that have been proved to have potential antitumour activity. However, the intracellular molecular targets of SLs and the underlying molecular mechanisms have not been well elucidated. Here, we report that F1012‐2, a novel SL active fraction, isolated from Eupatorium lindleyanum DC., can significantly inhibit the growth of triple‐negative breast cancer (TNBC) cells (MDA‐MB‐231 and MDA‐MB‐468) but has no obvious inhibitory effect on the growth of human mammary epithelial cells (MCF‐10A). The related mechanisms on cell growth inhibition of F1012‐2 were demonstrated by inducing apoptosis in a caspase‐dependent manner through the intrinsic pathway and extrinsic pathway. F1012‐2 could also activate autophagy in TNBC cells. Simultaneously, we found that F1012‐2‐induced apoptosis was enhanced by inhibition of autophagy. Furthermore, F1012‐2 could induce cell cycle arrest at G2/M phase with decreasing expression of cyclin B1, cdc2, and upregulating p21, p‐cdc2. Also, F1012‐2 activated Akt and p38 signalling pathways. In vivo, F1012‐2 exhibited a potential antitumour effect in MDA‐MB‐231 xenografts without apparent toxicity. Taken together, our results identified that F1012‐2 inhibited cell growth via multiple signalling pathways in vitro and in vivo. These data suggest that F1012‐2 may be a potential natural active fraction for the treatment of TNBC.     

Curcumin co-treatment ameliorates resistance to gefitinib in drug-resistant NCI-H1975 lung cancer cells

OBJECTIVE: To examine whether a combinative treatment with curcumin enhances the effects of the epidermal growth factor receptor-tyrosine kinase inhibitor(EGFR-TKI) gefitinib on cell proliferation, clonogenic capacity and apoptosis in the drug-resistant lung cancer cell line NCI-H1975, and further investigate the molecular mechanisms involved.METHODS: NCI-H1975 cells were treated with curcumin and gefitinib alone or in combination, and cell proliferation, clonogenic capacity and apoptosis were examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, clone forming experiments, and flow cytometry, respectively, while p38, extracellular regulated protein kinase(ERK)1/2, and protein kinase B(AKT)phosphorylation were examined using Western blotting.RESULTS: Compared with the effects of either agent alone, the combination of curcumin and gefitinib had a stronger suppressive effect on proliferation and the clonogenic capacity(P 0.05), and showed an increased ability to promote apoptosis(P 0.05) and reduce p38, ERK1/2, and AKT phosphorylation(P 0.05).CONCLUSION: Co-treatment of curcumin and gefitinib significantly improves the ability of gefitinib to inhibit cell proliferation, suppress the clonogenic capacity and enhance apoptosis in NCI-H1975 cells,and these effects are possibly mediated via a decrease in phosphorylation of proteins in downstream pathways of the epidermal growth factor receptor.     

Atractylenolide II induces G1 cell-cycle arrest and apoptosis in B16 melanoma cells

Ethnopharmacological relevance

Atractylenolide II (AT-II) is a sesquiterpene compound isolated from the dried rhizome of Atractylodes macrocephala (Baizhu in Chinese), which is traditionally prescribed for melanoma treatment by Chinese medicine practitioners. Our previous study showed that AT-II can inhibit B16 cells proliferation. Here we investigate the mechanistic basis for the anti-proliferative activity of AT-II in B16 melanoma cells.

Materials and methods

Cell viability was examined by MTT assay. Cell cycle distribution and apoptosis were determined by flow cytometry. Protein expression was determined by Western blotting.

Results

AT-II treatment for 48 h dose-dependently inhibited cell proliferation with an IC₅₀ of 82.3 μM, and induced G1 phase cell cycle arrest. Moreover, treatment with 75 μM AT-II induced apoptosis. These observations were associated with the decrease of the expression of Cdk2, phosphorylated-Akt, phosphorylated-ERK and Bcl-2, the increase of the expression of phosphorylated-p38, phosphorylated-p53, p21, p27, and activation of caspases-8, -9 and -3. In addition, a chemical inhibitor of p53, PFTα, significantly decreased AT-II-mediated growth inhibition and apoptosis.

Conclusions

We demonstrated that the G1-arresting and apoptotic effects of AT-II in B16 cells involve p38 activation as well as ERK and Akt inactivation, and the cytotoxic/apoptotic effects of AT-II are potentially p53 dependent. These findings provided chemical and pharmacological basis for the traditional application of Baizhu in melanoma treatment.     

Tanshinone IIA Induces Autophagic Cell Death via Activation of AMPK and ERK and Inhibition of mTOR and p70 S6K in KBM‐5 Leukemia Cells

Although tanshinone IIA (Tan IIA) from Salviae miltiorrhizae was known to induce apoptosis in various cancers, its underlying mechanism of autophagic cell death was not reported yet. Thus, in the present study, the molecular mechanism of autophagic cell death by Tan IIA was investigated in KBM‐5 leukemia cells. Tan IIA significantly increased the expression of microtubule‐associated protein light chain 3 (LC3) II as a hallmark of autophagy in western blotting and immunofluorescence staining. Tan IIA augmented the phosphorylation of adenosine monophosphate‐activated protein kinase (AMPK) and attenuated the phosphorylation of mammalian target of rapamycin (mTOR) and p70 S6K in a dose‐dependent manner. Conversely, autophagy inhibitor 3‐methyladenine partly reversed the cytotoxicity and the phosphorylation of AMPK, mTOR and p70 S6K induced by Tan IIA in KBM‐5 leukemia cells. In addition, Tan IIA dramatically activated the extracellular signal regulated kinase (ERK) signaling pathway including Raf, ERK and p90 RSK in a dose‐dependent and time‐dependent manner. Consistently, ERK inhibitor PD184352 suppressed LC3‐II activation induced by Tan IIA, whereas PD184352 and PD98059 did not affect poly (ADP‐ribose) polymerase cleavage and sub‐G1 accumulation induced by Tan IIA in KBM‐5 leukemia cells. Furthermore, Tan IIA could induce autophagy via LC3‐II activation in various cancer cells such as prostate (PC‐3), multiple myeloma (U266), lung (NCI‐H460), and breast (MDA‐MB‐231) cells. Overall, these findings suggest that Tan IIA induces autophagic cell death via activation of AMPK and ERK and inhibition of mTOR and p70 S6K in KBM‐5 cells as a potent natural compound for leukemia treatment. Copyright © 2013 John Wiley & Sons, Ltd.     

Dual inhibition of EGFR and MET by Echinatin retards cell growth and induces apoptosis of lung cancer cells sensitive or resistant to gefitinib

Patients with non‐small‐cell lung cancer (NSCLC) containing epidermal growth factor receptor (EGFR) amplification or sensitive mutations initially respond to tyrosine kinase inhibitor gefitinib; however, the treatment is less effective over time. Gefitinib resistance mechanisms include MET gene amplification. A therapeutic strategy targeting MET as well as EGFR can overcome resistance to gefitinib. In the present study we identified Echinatin (Ecn), a characteristic chalcone in licorice, which inhibited both EGFR and MET and strongly altered NSCLC cell growth. The antitumor efficacy of Ecn against gefitinib‐sensitive or –resistant NSCLC cells with EGFR mutations and MET amplification was confirmed by suppressing cell proliferation and anchorage‐independent colony growth. During the targeting of EGFR and MET, Ecn significantly blocked the kinase activity, which was validated with competitive ATP binding. Inhibition of EGFR and MET by Ecn decreases the phosphorylation of downstream target proteins ERBB3, AKT and ERK compared with total protein expression or control. Ecn induced the G2/M cell cycle arrest, and apoptosis via the intrinsic pathway of caspase‐dependent activation. Ecn induced ROS production and GRP78, CHOP, DR5 and DR4 expression as well as depolarized the mitochondria membrane potential. Therefore, our results suggest that Ecn is a promising therapeutic agent in NSCLC therapy.     

琼玉膏含药血清对化疗药物抑制肺腺癌细胞株GLC-82体外培养增效作用的实验研究

目的:观察琼玉膏含药血清对化疗药物抑制肺腺癌细胞株GLC-82体外培养增效作用。方法:使用细胞培养及计数方法绘制癌细胞株的生长曲线;用血清药理学、流式细胞等技术观察琼玉膏对肺腺癌细胞株GLC-82的细胞周期及凋亡的影响。结果:细胞形态学观察显示,化疗组与联合组对人肺癌细胞均有明显抑制作用,但联合组抑制作用更显著;细胞生长曲线显示,琼玉膏可明显加强顺氨氯铂(DDP)抑制癌细胞株的分裂;而且琼玉膏具有明显加强DDP延长癌细胞G1期(P<0.05)、降低s期(P<0.01)的作用,并可明显加强DDP诱发癌细胞的凋亡(P<0.01)。结论:琼玉膏有加强化疗药物抑制肺腺癌细胞分裂及诱发其凋亡作用。     

连翘苷经PI3K/Akt信号通路干预肾细胞癌的机制研究

目的探讨连翘苷抑制人肾细胞腺癌786-0细胞生长、迁移及侵袭的作用机制。方法体外培养786-0细胞,在其中加入不同浓度连翘苷进行干预。MTT法检测细胞生存活力,AO/EB法检测细胞凋亡,划痕实验与Transwell实验分别考察细胞迁移、侵袭能力。Western blotting法检测细胞内PI3K、p-PI3K、Akt、p-Akt、FOXO3a、p-FOXO3a、p21、p27、Fasl、Bim、MMP-2及MMP-9蛋白表达情况。结果连翘苷可有效抑制肾癌细胞生长、促进凋亡,干扰细胞周期,上调p21、p27、Fasl及Bim表达水平,与对照组比较差异明显(P0.05、0.01);与对照组比较,不同浓度连翘苷可明显抑制肾癌细胞迁移与侵袭,减少MMP-2、MMP-9合成(P0.05、0.01);同时连翘苷能明显抑制PI3K、Akt、FOXO3a磷酸化(P0.05、0.01),且呈浓度依赖性。结论连翘苷可通过PI3K/Akt信号通路调控786-0细胞凋亡与细胞周期、抑制肾癌细胞生长;同时连翘苷经PI3K/Akt通路可有效削弱肾癌细胞迁移与侵袭能力。     

�˲ζ����յ��˸ΰ�BEL-7402ϸ���������ü�����

??OBJECTIVE To investigate the apoptosis of the auction and mechanism of the hepatoma BEL-7402 cells induced by the ginseng polysaccharides(GPS). METHODS The hematoma cells BEL-7402 were incubated with GPS, cell viability was measured by CCK8, cell cycle distribution was assessed by fluorescence-activated cell sorting(FACS), the cell morphological changes were traced with TUNEL and scanning electron microscope, TNFR1, BCL-2 and Bax protein expression and ERK phosphorylation are tested by Western blot. RESULTS CCK8 results showed that GPS inhibited cells growth of BEL-7402 in dose-dependent and time-dependent. Flow cytometry found that S phase arrest was increased upon GPS concentrations. Obviously apoptotic sub-g peak was also found. And the peak was gradually enhanced with the concentration increased. TUNEL staining and SEM results showed that GPS led to significant cell morphology changes in hepatoma cells BEL-7402.And the apoptosis effect was increased upon the GPS concentrations. Western blot showed that level of apoptosis related protein Bax was increased, the expression of apoptosis-antagonizing protein Bcl-2 was decreased and the expression of death receptor TNFR1 appeared with GPS concentrations increased gradually,raise ERK phosphorylation. CONCLUSION GPS can induce apoptosis of hepatoma cells BEL-7402 by the mitochondrial pathway and the death receptor dependent pathway and ERK pathway.     

Antiproliferative and proapoptotic actions of okra pectin on B16F10 melanoma cells

The proliferation and apoptosis of metastatic melanoma cells are often abnormal. We have evaluated the action of a pectic rhamnogalacturonan obtained by hot buffer extraction of okra pods (okra RG‐I) on melanoma cell growth and survival in vitro. We added okra RG‐I containing an almost pure RG‐I carrying very short galactan side chains to 2D (on tissue culture polystyrene, tPS) and 3D (on poly(2‐hydroxyethylmethacrylate), polyHEMA) cultures of highly metastatic B16F10 mouse melanoma cells. We then analyzed cell morphology, proliferation index, apoptosis, cell cycle progression and the expression of adhesion molecules. Immunostaining and western blotting were used to assay galectin‐3 (Gal‐3) protein. Incubation with okra RG‐I altered the morphology of B16F10 cells and significantly reduced their proliferation on both tPS and polyHEMA. The cell cycle was arrested in G2/M, and apoptosis was induced, particularly in cells on polyHEMA. The expression of N‐cadherin and α5 integrin subunit was reduced and that of the multifunctional carbohydrate‐binding protein, Gal‐3, at the cell membrane increased. These findings suggest that okra RG‐I induces apoptosis in melanoma cells by interacting with Gal‐3. As these interactions might open the way to new melanoma therapies, the next step will be to determine just how they occur. Copyright © 2009 John Wiley & Sons, Ltd.     

Paris saponin VII induces cell cycle arrest and apoptosis by regulating Akt/MAPK pathway and inhibition of P‐glycoprotein in K562/ADR cells

Paris saponinVII (PSVII) is a steroidal saponin isolated from the roots and rhizomes of Trillium tschonoskii Maxim. We found that PSVII could inhibit the growth of adriamycin‐resistant human leukemia cells (K562/ADR) in a dose‐dependent manner. Furthermore, the molecular mechanism underlying the cytotoxicity and downregulation of P‐glycoprotein (P‐gp) expression by PSVII was clarified. PSVII significantly suppressed cell proliferation by cell cycle arrest in the G0/G1 phase, which was associated with an obvious decrease in cyclin B1/D1 and CDK2/4/6 protein expression. Moreover, PSVII could attenuate mitochondrial membrane potential, increase the expression of apoptosis‐related proteins, such as Bax and cytochrome c, and decrease the protein expression levels of Bcl‐2, caspase‐9, caspase‐3, PARP‐1, and p‐Akt. We also found that JNK, ERK1/2, and p38 were regulated by PSVII in K562/ADR cells. And further studies indicated that the decrease in the reactive oxygen species level inhibited intrinsic P‐gp expression. Therefore, PSVII‐induced apoptosis in K562/ADR cells was associated with Akt/MAPK and the inhibition of P‐gp. In addition, PSVII induced a robust autophagy in K562/ADR cells as demonstrated by the degradation of LC3‐I. These results provide a biochemical basis for possible clinical applications of PSVII in the treatment of leukemia.     

Synergistic Effect of TPD7 and Berberine against Leukemia Jurkat Cell Growth through Regulating Ephrin‐B2 Signaling

TPD7, a novel biphenyl urea taspine derivative, and berberine have presented inhibition on VEGFR2 that can be regulated by ephrin‐B2 reverse signaling through interactions with the PDZ domain. The purpose of this study is to investigate the inhibitory effect of the combination of TPD7 and berberine (TAB) on T‐cell acute lymphoblastic leukemia cell growth. TPD7 and berberine together synergistically inhibited the proliferation of Jurkat cells. Also, the combination of TAB induced G₁‐phase cell‐cycle arrest by downregulating the level of cyclin D1, cyclin E, and CDC2. Furthermore, the combination of TAB significantly enhanced apoptosis in Jurkat cells, and the apoptosis most likely resulted from the modulation of the level of Bcl‐2 family members. Most importantly, the concomitant treatment simultaneously regulated the ephrin‐B2 and VEGFR2 signaling, as well as modulated the MEK/ERK and PTEN/PI3K/AKT/mTOR signaling. Therefore, the combination treatment of TAB may be a promising therapeutic method in treating T‐cell acute lymphoblastic leukemia. Copyright © 2017 John Wiley & Sons, Ltd.     

Lapachol is a novel ribosomal protein S6 kinase 2 inhibitor that suppresses growth and induces intrinsic apoptosis in esophageal squamous cell carcinoma cells

Lapachol is a 1,4‐naphthoquinone that is isolated from the Bignoniaceae family. It has been reported to exert anti‐inflammatory, antibacterial, and anticancer activities. However, the anticancer activity of lapachol and its molecular mechanisms against esophageal squamous cell carcinoma (ESCC) cells have not been fully investigated. Herein, we report that lapachol is a novel ribosomal protein S6 kinase 2 (RSK2) inhibitor that suppresses growth and induces intrinsic apoptosis in ESCC cells. We found that lapachol strongly attenuates downstream signaling molecules of RSK2 in ESCC cells and also directly inhibits RSK2 activity in vitro. The RSK protein is highly activated in ESCC cells and knockdown of RSK2 significantly suppresses anchorage‐dependent and anchorage‐independent growth of ESCC cells. Additionally, lapachol inhibits anchorage‐dependent and anchorage‐independent growth of ESCC cells, and the inhibition of cell growth by lapachol is dependent on the expression of RSK2. We also found that lapachol induces mitochondria‐mediated cellular apoptosis by activating caspases‐3, ‐7, and PARP, inducing the expression of cytochrome c and BAX by inhibiting downstream molecules of RSK2. Overall, lapachol is a potent RSK2 inhibitor that might be used for chemotherapy against ESCC.     

Apoptosis Induction by 13‐Acetoxyrolandrolide through the Mitochondrial Intrinsic Pathway

The aim of this study was to evaluate the mechanisms of cytotoxicity of the sesquiterpene lactone 13‐acetoxyrolandrolide, a nuclear factor kappa B (NF‐κB) inhibitor that was previously isolated from Rolandra fruticosa. The effects associated with the inhibition of the NF‐κB pathway included dose‐dependent inhibition of the NF‐κB subunit p65 (RelA) and inhibition of upstream mediators IKKβ and oncogenic Kirsten rat sarcoma (K‐Ras). The inhibitory concentration of 13‐acetoxyrolandrolide on K‐Ras was 7.7 µm . The downstream effects of the inhibition of NF‐κB activation were also investigated in vitro. After 24 h of treatment with 13‐acetoxyrolandrolide, the mitochondrial transmembrane potential was depolarized in human colon cancer (HT‐29) cells. The mitochondrial oxidative phosphorylation was also negatively affected, and reduced levels of nicotinamine adenine dinucleotide phosphate (NAD(P)H) were detected after 2 h of 13‐acetoxyrolandrolide exposure. Furthermore, the expression of the pro‐apoptotic protein caspase‐3 increased in a concentration‐dependent manner. Cell flow cytometry showed that 13‐acetoxyrolandrolide induced cell cycle arrest at G₁, indicating that the treated cells had undergone caspase‐3‐mediated apoptosis, indicating negative effects on cancer cell proliferation. These results suggest that 13‐acetoxyrolandrolide inhibits NF‐κB and K‐Ras and promotes cell death mediated through the mitochondrial apoptotic pathway. Copyright © 2013 John Wiley & Sons, Ltd.     

Protolichesterinic Acid,Isolated from the Lichen Cetraria islandica,Reduces LRRC8A Expression and Volume‐Sensitive Release of Organic Osmolytes in Human Lung Epithelial Cancer Cells

We have tested the effect of protolichesterinic acid (PA) on the activity of the volume‐sensitive release pathway for the organic osmolyte taurine (VSOAC) and the expression of the leucine‐rich‐repeat‐channel 8A (LRRC8A) protein, which constitutes an essential VSOAC component. Exposing human lung cancer cells (A549) to PA (20 µg/mL, 24 h) reduces LRRC8A protein expression by 25% and taurine release following osmotic cell swelling (320 → 200 mOsm) by 60%. C75 (20 µg/mL, 24 h), a γ‐lactone with a C8 carbon fatty acid chain, reduces VSOAC activity by 30%, i.e. less than PA. Stearic acid (20 µg/mL, 24 h) has no effect on VSOAC. Hence, length of PA's fatty acid chain adds to γ‐lactone's inhibitory action. 5‐Lipoxygenase (5‐LO) activity is essential for swelling‐induced activation of VSOAC. PA has no effect on cellular concentration of leukotrienes (5‐HETE/LTB₄) under hypotonic conditions, excluding that PA mediated inhibition of VSOAC involves 5‐LO inhibition. A549 cells exposed to the chemotherapeutic drug cisplatin (10 μM, 24 h) reveal signs of apoptosis, i.e. 25% reduction in cell viability as well as 1.3‐, 1.5‐ and 3.3‐fold increase in the expression of LRRC8A, Bax (regulator of apoptosis) and p21 (regulator of cell cycle progression), respectively. PA reduces cell viability by 30% but has no effect on p21/Bax expression. This excludes PA as a pro‐apoptotic drug in A549 cells. Copyright © 2015 John Wiley & Sons, Ltd.     

菩人丹对高糖诱导的INS-1细胞凋亡、BAD和FOXO1蛋白表达的影响

目的: 观察菩人丹(Puren Dan, PRD)对高糖诱导的INS-1细胞凋亡、Bal-xl/Bcl-2相关死亡促进因子(BAD)和叉头框转录因子O亚族1(FOXO1)蛋白表达的影响,探讨PRD恢复INS-1细胞分泌功能的相关分子机制。 方法: 采用血清药理学方法制备菩人丹含药血清,建立高浓度葡萄糖(33.3 mmol·L^-1)诱导INS-1细胞损伤模型,以菩人丹药物血清干预24 h;实验分为5组,即正常对照组(control组)、高糖模型组(HG组)、菩人丹药物血清高剂量组(H-PRD组)、菩人丹药物血清低剂量组(L-PRD组)和二甲双胍药物血清对照组(MF组);采用CCK8试剂盒检测细胞活力,流式Annexin V-FITC/PI双染法考察细胞凋亡水平,Western blotting assay检测BAD和FOXO1蛋白表达水平及其磷酸化水平。 结果: 与正常组比较高糖能够显著降低INS-1细胞活力、诱导细胞凋亡、降低INS-1细胞BAD,FOXO1丝氨酸磷酸化水平;与模型组比较菩人丹含药血清(终体积分数10%)则能抑制INS-1细胞凋亡,增加细胞活力,下调FOXO1表达,促进BAD和FOXO1丝氨酸磷酸化。 结论: 菩人丹药物血清能够减少高糖诱导的INS-1细胞凋亡,这种作用与上调BAD和FOXO1蛋白丝氨酸磷酸化水平直接相关。     

Matrine triggers colon cancer cell apoptosis and G0/G1 cell cycle arrest via mediation of microRNA‐22

Matrine (MAT) is an alkaloid in the dried roots of Sophora flavescens. The antitumor activity has been testified in colon cancer. Howbeit, the latent mechanism is still indistinct. The research probed the antitumor mechanism of MAT in colon cancer cells. MAT (0.25, 0.5, 0.75, 1, and 1.25 mM) was utilized to stimulate SW480 and SW620 cells for 24, 48, and 72 hr. Cell viability, apoptosis, cell cycle, and the correlative proteins were assessed via Cell Counting Kit‐8, flow cytometry, and Western blot. microRNA‐22 (miR‐22) in MAT‐treated or miR‐22‐silenced cells was estimated via real‐time quantitative polymerase chain reaction. The functions of miR‐22 inhibition were reassessed. Western blot was conducted for quantifying β‐catenin, MEK, and ERK. Luciferase reporter assay was done for confirming the targeting relationship between miR‐22 and ERBB3 or MECOM. MAT prohibited cell viability, accelerated apoptosis, and triggered cells cycle stagnation at G0/G1 phase. Additionally, miR‐22 was elevated by MAT; meanwhile, the influences of MAT were all inverted by miR‐22 inhibitor. MAT enhanced the expression of miR‐22, thereby obstructing Wnt/β‐catenin and MEK/ERK pathways. miR‐22 had a potential to target mRNA 3'UTR of ERBB3 and MECOM. These discoveries manifested that MAT could evoke colon cancer cell apoptosis and G0/G1 cell cycle arrest via elevating miR‐22.