Expression and Emmprin Regulated Function of Aquaporin1 in SMMC7221 Cells

Herein lie the crosstalk and regulation between AQP1 and Emmprin in SMMC7221 cells by means of siRNA technology and deglycosylation method. Firstly, HAQP1, rather than hAQP3, was selectively upregulated in SMMC7221 cells by FBS, flollowed by the upregulated expression of Emmprin. Emmprin gene silencing caused a remarkable change in the expression of AQP1 gene, just like its downstream gene, MMP9, meanwhile the water permeability and cell migration were also descended prominently. Furthermore, when treated with tunicamycin, Emmprin was deglycosylated, which made the expression of AQP1 significantly declined, followed by remarkably decreased cell membrane water permeability and cell migration. Taken together, all the data indicates the expression level and the modification of Emmprin by glycosylation are the key factors in regulating the expression of AQP1.     

Synthesis and biological evaluation of picroside derivatives as hepatoprotective agents

Picrorhizae Rhizoma as a hepatoprotective herb, has been applied for thousands of years, and picroside was proved to be its active constituent. In this study, twelve derivatives of picroside were synthesized and the hepatoprotective activity of the derivatives was evaluated on SMMC-7721 cells. Six out of the derivatives had shown a better protective effect on H₂O₂-induced SMMC-7221 cells than picroside, and the activity of two derivatives (2 and 4) was stronger than that of the reference compound, silybin. Compound 2 shown the strongest protective effect (EC₅₀?=?6.064?±?1.295?μM).     

苯乙酸对肝癌细胞系SMMC-7721的增殖抑制作用及与RNA编辑酶ADAR1表达的相关性

为探讨苯乙酸(PA)对肝癌细胞系SMMC-7721的增殖抑制作用及其与RNA编辑酶ADAR1表达的相关性, 应用细胞计数及MTT法检测了不同浓度(0.5, 1.0, 2.0和4.0 mmol/L)PA对肝癌细胞系SMMC-7721的增殖抑制作用, 通过流式细胞术(FCM)分析了各细胞周期的细胞百分比, 应用半定量逆转录-聚合酶链式反应(RT-PCR)及免疫印迹杂交分析使用不同浓度(0.5, 1.0, 2.0 mmol/L)PA作用后肝癌细胞系SMMC-7721中RNA编辑酶ADAR1 mRNA及蛋白表达的变化. 结果表明, 肝癌细胞系SMMC-7721经不同浓度PA作用后, 增殖抑制率随作用时间延长及PA浓度增加而明显提高(P<0.05), 但2.0和4.0 mmol/L PA作用72 h后组间差异比较无统计学意义(P>0.05). 肝癌细胞系SMMC-7721中RNA编辑酶ADAR1 mRNA及蛋白表达随PA浓度增加而明显降低(P<0.05). 通过沉默SMMC-7721细胞中ADAR1的表达发现, ADAR1表达下调可有效抑制肝癌细胞增殖. 结果表明, PA可阻抑肝癌细胞系SMMC-7721细胞增殖, 且存在时间及剂量的依赖性, 作用机制与PA下调ADAR1表达相关.     

Design,synthesis, and biological evaluation of sorafenib derivatives containing indole (ketone) semicarbazide analogs as antitumor agents

A series of new sorafenib derivatives was designed and synthesized. The antiproliferative activity of the synthesized compounds against human lung cancer cell (A549), human pancreatic cancer cell (PC-3), human leukemia cell (K562), and human hepatoma cell (SMMC-7721) was evaluated by MTT assay. The results revealed that several compounds displayed more significant antitumor activities than commercial anticancer agent sorafenib against SMMC-7721. In addition, compounds 7a , 7g , 7l , 7m , and 7p represented obvious growth inhibition with IC₅₀ values of 1-9 μM against four cancer cell lines, demonstrating more predominant activities against cancer cells as compared to sorafenib. Furthermore, some structure-activity relationships have also been established. Compounds containing indole and benzene ring substituted by halogen showed better activity than sorafenib. Wound healing assay suggested that cells would be targeted on their migratory capacity by 7g , potentially affecting the migration activity of these tumors. The effects of A549 and PC-3 cell apoptosis induced by compound 7g were significantly increased compared with sorafenib. Importantly, the result of western blot assay showed that 7g inhibited cell growth by suppressing the activity of EGFR, especially the expression of p-EGFR (Tyr1068).     

Apoptosis Induced by Aqueous Extracts of Crocodile Bile in Human Heptacarcinoma SMMC-7721

In the present study, effects of aqueous extracts from Crocodylus siamensis bile (AE-CB) on SMMC-7721 cell growth, cell cycle, and apoptosis were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, inverted microscopy, fluorescence microscopy, propidium iodide (PI) single- and fluorescein isothiocyanate (FITC)/PI double-staining flow cytometry, and western blotting. Our data have revealed that AE-CB significantly inhibited the growth of SMMC-7721 cell and arrested cell cycle at G0/G1 phase. SMMC-7721 cells showed typical apoptotic morphological changes after treated with AE-CB for 48 h. Cell death assay indicated that SMMC-7721 cells underwent apoptosis in a dose-dependent manner induced by AE-CB. In addition, AE-CB treatment could downregulate the protein level of Bcl-2 and upregulate the Bax, leading to the increase in the ratio of Bax to Bcl-2 in SMMC-7721 cells. Meanwhile, it was observed that the expression of Survivin and c-Myc decreased, but the expression of P53 increased. All these events were associated with increase of reactive oxygen species. The data indicated that mitochondrial pathway might play an important role in bile extract-induced apoptosis in SMMC-7721 cells. These results provide significant insight into the anticarcinogenic action of bile extract on SMMC-7721 cells.     

Anti-migration effects of Gekko sulfated glycopeptide on human hepatoma SMMC-7721 cells

Gekko swinhonis Guenther has been used as an anti-cancer drug in traditional Chinese medicine for hundreds of years. Previous studies showed that the Gekko sulfated polysaccharide-protein complex suppressed the proliferation and migration of hepatoma cells. Gekko sulfated glycopeptide α was obtained from Gekko sulfated polysaccharide-protein complex using papain hydrolysis. Gekko sulfated glycopeptide α inhibited the proliferation and migration of SMMC-7721 cells. The secretion of IL-8 and the concentration of intracellular calcium were decreased after Gekko sulfated glycopeptide α exposure. SMMC-7721 cells in the control group showed abnormal features, with a polygonal shape, whereas this changed to a spindle shape after the treatment with Gekko sulfated glycopeptide α. Actin ?laments were distributed diffusely along the cell membrane in control cells, whereas those were polymerized and preferentially accumulated in the cytoplasm of treated cells. Microtubules distributed in the cytoplasm of untreated cells were located diffusely whereas those in treated cells were polymerized. Therefore, Gekko sulfated glycopeptide α inhibit the migration of hepatoma cells via reducing the secretion of IL-8 and the concentration of intracellular calcium, as well as regulating the reorganization of cytoskeleton.     

Effects of oxymatrine on proliferation and apoptosis in human hepatoma cells

Oxymatrine, a natural quinolizidine alkaloid, has been known having cytotoxic and chemopreventive effects on various cancer cells. To investigate the possible mechanism of oxymatrine's role on cancer cells, in the present study, we examined further the effects of oxymatrine on the growth, proliferation, apoptosis and expression of bcl-2 and p53 gene in human hepatoma SMMC-7721 cells in vitro. Our results show that oxymatrine notably inhibits the growth and proliferation of SMMC-7721 cells and it present a dose-dependence and time-dependence manner within definite reacting dose and time. Oxymatrine block SMMC-7721 cells in G2/M and S phase; prevent cells entering into G0/G1 phase. It results in an obvious accumulation of G2/M and S phase cells while decrease of G0/G1 phase cells. Oxymatrine induce apoptosis of SMMC-7721 cells and apoptotic rate amount to about 60% after treatment with 1.0 mg/ml oxymatrine for 48 h. We also find that oxymatrine down-regulate expression of bcl-2 gene while up-regulate expression of p53 gene. These results demonstrate that oxymatrine inhibit the proliferation and induce apoptosis of human hepatoma SMMC-7721 cells, and suggest that this effect was mediated probably by a significant cell cycle blockage in G2/M and S phase, down-regulation of bcl-2 and up-regulation of p53.     

麦斛细胞毒活性成分研究

利用硅胶、聚酰胺柱层析等分离方法, 从麦斛全草的乙醚萃取物中分离得到4个化合物, 经化学方法和光谱分析(UV, IR, NMR, MS)鉴定为3,4,7-三羟基-2-甲氧基菲(1), 2,5-二羟基-4-甲氧基菲(2), eugenitin (3)和2,5-二羟基-4-甲氧基-9,10-二氢菲(4), 其中1为新化合物. 采用MTT法分别测得化合物1和2对体外培养的人急性髓性白血病细胞和人肝癌细胞增殖的抑制活性, 并分别考察了增敏剂葛根素对化合物1和2增殖活性的协同作用.     

新型双氢青蒿素哌嗪-脂肪族酰胺类化合物的合成及其抗癌活性

以双氢青蒿素（DHA）为原料,与草酰氯和哌嗪经“一锅”法制得DHA哌嗪衍生物（2）; 2与脂肪族酰氯经酰化反应合成了6个新型的双氢青蒿素哌嗪-酰胺类衍生物（4a～4f）,其结构经¹H NMR, ¹³C NMR,IR和HR-ESI-MS进行表征。以四甲基偶氮唑盐比色法（MTT法）初步研究了4a～4f对人肝癌细胞株SMMC-7721的抑制活性。结果表明,4a～4f显著抑制SMMC-7721的增殖,并诱导其凋亡。其中,双氢青蒿素哌嗪-氯乙酰胺（4c）的活性最好,IC₅₀为0.05 μM,优于青蒿素（IC₅₀ 0.53 μM）和DHA（IC₅₀0.52 μM）。     

Krüppel-like factor 4 mediates lysophosphatidic acid-stimulated migration and proliferation of PC3M prostate cancer cells

Prostate cancer is the most frequently diagnosed malignancy and the second leading cause of cancer mortality among men in the United States. Accumulating evidence suggests that lysophosphatidic acid (LPA) serves as an autocrine/paracrine mediator to affect initiation, progression and metastasis of prostate cancer. In the current study, we demonstrate that LPA stimulates migration and proliferation of highly metastatic human prostate cancer, PC-3M-luc-C6 cells. LPA-induced migration of PC-3M-luc-C6 cells was abrogated by pretreatment of PC-3M-luc-C6 cells with the LPA receptor 1/3 inhibitor Ki16425 or small interfering RNA (siRNA)-mediated silencing of endogenous LPA receptor 1, implicating a key role of the LPA-LPA receptor 1 signaling axis in migration of PC-3M-luc-C6 cells. In addition, LPA treatment resulted in augmented expression levels of Krüppel-like factor 4 (KLF4), and siRNA or short-hairpin RNA (shRNA)-mediated silencing of KLF4 expression resulted in the abolishment of LPA-stimulated migration and proliferation of PC-3M-luc-C6 cells. shRNA-mediated silencing of KLF4 expression resulted in the inhibition of in vivo growth of PC-3M-luc-C6 cells in a xenograft transplantation animal model. Taken together, these results suggest a key role of LPA-induced KLF4 expression in cell migration and proliferation of prostate cancer cells in vitro and in vivo.     

Trichiliasinenoids A–C,three 6,7-secomexicanolide limonoids with a 7,29-linkage from Trichilia sinensis

Trichiliasinenoids A–C (1–3), three new limonoids with an unprecedented C-29-C-7 connecting carbon skeleton formed by migration of C-7 from C-6 to C-29 of a mexicanolide-type limonoid precursor were isolated from the leaves and twigs of Trichilia sinensis. Their structures were assigned by spectroscopic analysis, and the absolute configurations were determined by X-ray crystallography (1) and ECD calculation (2 and 3). A possible biosynthetic pathway of 1 was also proposed. Compound 2 exhibited moderate cytotoxicity against HL-60 cells and weak cytotoxicity against SMMC-7721cells.     

千里光中四个新倍半萜的结构

从植物千里光中分得四个新艾里莫芬烷型倍半萜,经仔细的波谱分析和单晶X射线衍射,研究它们的结构分别为为：7β,11-环氧-9α,10α-环氧-8-羰基艾里莫芬烷(1),8-11-过氧-9α,10α-环氧-6-烯-8β-羟基艾里莫芬烷(2),7(11)-烯-9α,10α-环氧-8-羰基艾里莫芬烷(3),6-烯-9α,10α-环氧-11-羟基-8-羰基艾里莫芬烷(4)。活性筛选实验表明化合物1具有促进癌细胞生长作用,化合物2和3具有较好的抗癌活性。     

3,4,5-三羟基苯甲酸通过线粒体途径诱导人肝癌细胞SMMC-7721的凋亡

采用四甲基偶氮唑盐(MTT)观察3,4,5-三羟基苯甲酸(TBA)对人肝癌SMMC-7721细胞的增殖抑制作用; 通过流式细胞仪检测细胞凋亡、细胞内活性氧(ROS)及线粒体膜电位的变化, 用比色法测定Caspase-9和Caspase-3蛋白活性, 探讨TBA 诱导SMMC-7721细胞凋亡的分子机制. 研究结果表明, TBA对SMMC-7721细胞生长具有显著的抑制作用并诱导其凋亡, 诱导SMMC-7721细胞凋亡作用可能通过线粒体信号传导通路实现.     

Synthesis of Some Allyl-β-Cyclodextrin Derivatives and their Properties as Capillary GC Stationary Phases

Summary Three -Cyclodextrin derivatives (-CDs) were synthesized by substituting the 2,6-OH groups of -CD with allyl groups and the 3-OH groups with three different acyl groups (valeryl, heptanonyl, octanoyl). The chromatographic properties of these -CDs as stationary phases for capillary gas chromatography (CGC) were studied. The test results showed that the three -CDs possessed good coating properties and that the capillary columns coated with them exhibited high column efficiency. These -CDs can separate not only the disubstituted benzene isomers but also some racemic compounds.     

Components of Volatile Oil from Water-caltrop and Their Anti-tumor Effect in vitro

The volatile oil was extracted from water caltrop by steam distillation; it was then analyzed by GC-MS to obtain 16 components, 8 of which were identified. Apocynin was the most abundant one, accounting for 81.41% of the total oil. The in vitro inhibitory effects of the volatile oil on SMMC-7721, MCF-7, Hela, HL-60 cells, and human peripheral blood mononuclear celIs(PBMC) were investigated via the MTT method. The morphological changes of the tumor cells were observed and the apoptosis of HL-60 cells was detected by flow cytometry. The proliferation of the tumor cells could be significantly inhibited and the apoptosis of HL-60 cells could be induced by the volatile oil. The proliferation inhibition effect of the volatile oil on HL-60 tumor cells and the induction of the apoptosis of HL-60 cells had dose-dependent feature.     

Essential oil of Cephalotaxus griffithii needle inhibits proliferation and migration of human cervical cancer cells: involvement of mitochondria-initiated and death receptor-mediated apoptosis pathways

This study was conducted to determine the effect of Cephalotaxus griffithii needle essential oil (CGNO) on proliferation and migration of human cervical cancer (HCC) cells. CGNO treatment decreased the viability of all the tested HCC (HeLa, ME-180 and SiHa) cells. Morphological and DNA fragmentation analysis of CGNO-treated HeLa cells indicated the involvement of apoptosis in inducing HCC cell death. CGNO increased mitochondrial membrane depolarisation and upregulated the expression of caspase-9, caspase-8, caspase-3 and cleaved-PARP. The activity of caspase-8 and caspase-9 was also significantly increased. Wound healing and transwell migration assay demonstrated that CGNO significantly inhibited the migration of HeLa cells to close a scratched wound and also inhibited their migration through filter towards a chemotactic stimulus. Taken together, these results indicated that CGNO inhibited the proliferation and migration of HCC cells. Of note, CGNO induced HeLa cell death through mitochondria-initiated and death receptor-mediated apoptosis pathway.     

Camptothecin-20(s)-O-[N-(3'α,12'α-dihydroxy-24'-carbonyl-5'β-cholan)]-lysine, a novel camptothecin analogue, induces apoptosis towards hepatocellular carcinoma SMMC-7721 cells

Camptothecin-20(s)-O-[N-(3'α,12'α-dihydroxy-24'-carbonyl-5'β-cholan)]-lysine (B2) is a novel camptothecin analogue. Our previous study had shown that it displayed higher cytoxicity activity towards hepatocellular carcinoma SMMC-7721 cells than camptothecin (CPT) in vitro. In this paper, the underlying mechanism of anti-proliferation of B2 towards SMMC-7721 cells was further examined. Cell growth inhibition of B2 was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; morphological changes were observed under Laser Scanning Confocal Microscope (LSCM); cell cycle distribution, apoptotic population, changes in mitochondrial membrane potential, intracellular calcium concentration and reactive oxygen species (ROS) production were determined by flow cytometry (FCM). Activities of caspase-3 and caspase-9 were measured, and the expression level of Bcl-2 and Bax proteins were analyzed by Western blot. The results suggested that B2 inhibited SMMC-7721 cell growth by causing cell cycle arrest at the S and G2/M phases, and induced apoptosis involving a mitochondrial pathway. B2 appears to cause a high induction of apoptosis on SMMC-7721 cells in vitro, which suggests it might be a potential drug for cancer therapy.     

Two new epimeric pairs of iridoid from mangrove plant Scyphiphora hydrophyllacea

Two new epimeric pairs of iridoid scyphiphin A_1(1a),A_2(1b) and scyphiphin B_1(2a),B_2(2b) were isolated from Scyphiphora hydrophyllacea Gaertn.F.Their structures were elucidated by spectroscopic methods.The mixture of scyphiphin B_1 and scyphiphin B_2 showed moderate cytotoxicity against human hepatoma SMMC-7721 cell line in vitro by MTT method.