Resveratrol inhibits proliferation, invasion and cell cycle of hepatobiliary carcinoma SMMC-7721 cells by downregulating the expression of PRMT5 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究白藜芦醇（Res）通过调控PRMT5表达对肝胆管癌SMMC-7721细胞增殖、侵袭、细胞周期的影响及其机制。方法：常规培养正常肝细胞LO2和SMMC-7721细胞，用0、20、40、80 μmol/L的Res进行处理，用qPCR法、MTT法、Transwell实验、流式细胞术和WB法分别检测Res处理后PRMT5 mRNA在LO2和SMMC-7721细胞中的表达，Res对SMMC-7721细胞增殖能力、侵袭能力、细胞周期和凋亡，以及PRMT5、cyclin D1和cyclin E1蛋白表达的影响。结果：PRMT5在SMMC-7721细胞中呈高表达（P<0.01）；20、40、80 μmol/L Res均能明显抑制PRMT5 mRNA和蛋白在SMMC-7721细胞中的表达（均P<0.01），抑制SMMC-7721细胞的增殖能力（P<0.01）和侵袭能力（P<0.05），阻滞SMMC-7721细胞周期于G0/G1期并促进其凋亡（P<0.01），明显抑制SMMC-7721细胞中周期蛋白cyclin D1、cyclin E1蛋白的表达（P<0.01）。结论：PRMT5在SMMC7721细胞中呈高表达，Res可有效抑制SMMC-7721细胞的增殖和侵袭能力并诱导其凋亡，其机制可能与抑制PRMT5表达相关。

Effect of Stemona tuberosa Alkaloids on Apoptosis of SMMC-7721 Cells and Expression of Bcl-2， Bax， and Cleaved Caspase-3 / 中国实验方剂学杂志

Objective:To investigate the effect of<italic> Stemona tuberosa</italic> alkaloids on the apoptosis of human hepatoma SMMC-7721 cells and the expression of apoptosis-related proteins including B lymphocytoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）. Method:SMMC-7721 cells were routinely cultured， passaged， and treated with various concentrations （50， 75， 112， 167， and 250 mg·L^-1） of <italic>S. tuberosa </italic>alkaloids， while those in the blank control group were only treated with 10% fetal bovine serum. The cell proliferation was determined by tetrazolium bromide （MTT） colorimetry and colony assay and the cell apoptosis by Hoechst 33258 staining. The protein expression levels of Bcl-2， Bax， and cleaved Caspase-3 were detected by Western blot. Result:<italic>S. tuberosa</italic> alkaloids inhibited the proliferation of SMMC-7721 cells， and the inhibition rate was significantly increased in comparison with that in the blank control group （<italic>P</italic><0.01）， with the half maximal inhibitory concentrations （IC₅₀） at 24 h， 48 h， and 72 h being （173.36±8.75）， （112.14±16.50）， and （96.41±2.60）mg·L^-1， respectively. The cell colony-inhibitory activity was significantly increased in a dose-dependent manner （<italic>P</italic><0.01）. Compared with the blank control group， <italic>S. tuberosa</italic> alkaloids promoted the apoptosis of SMMC-7721 cells， manifested as increased number of apoptotic cells and elevated apoptotic rate （<italic>P</italic><0.01）. The typical morphological changes such as brightly blue-fluorescent condensed nuclei， cytoplasmic shrinking， and karyopyknosis were found under the upright fluorescence microscope. As revealed by comparison with the blank control group， the expression of Bcl-2 was significantly down-regulated （<italic>P</italic><0.01）， while the protein expression levels of pro-apoptotic protein Bax and cleaved Caspase-3 in the 75， 112， 167， and 250 mg·L^-1 <italic>S. tuberosa</italic> alkaloids groups were significantly up-regulated （<italic>P</italic><0.01）. Conclusion:<italic>S. tuberosa </italic>alkaloids inhibit the proliferation of SMMC-7721 cells and promote their apoptosis possibly by inhibiting Bcl-2 protein expression and promoting Bax and cleaved Caspase-3 protein expression.

Screening of prognosis-related enhancer RNAs and their corresponding target genes in patients with hepatocellular carcinoma and its prognostic significance / 中国肿瘤生物治疗杂志

@#[摘 要] 目的： 筛选与肝细胞癌（hepatocellular carcinoma，HCC）预后相关的增强子RNA（enhancer RNA，eRNA）及其对应的靶基因，并探究其调控作用及功能。方法：通过UCSC数据库下载HCC的转录本数据和临床病例数据，提取eRNA及其预测的相应靶基因表达矩阵，用Kaplan-Meier和Spearman相关性分析方法筛选HCC预后相关的eRNA和靶基因，并采用最小化绝对收缩和选择算子回归分析及多因素Cox多元回归分析构建HCC预后风险评分模型。设计合成针对筛选所获关键eRNA AP003469.2的小干扰RNA并转染肝癌SMMC-7721细胞，采用RT-PCR方法验证敲减效率，qPCR法检测AP003469.2靶基因YWHAZ的表达水平，CCK-8法检测转染后SMMC-7721细胞的增殖情况。结果：筛选获得4个与预后相关基因，包括两个eRNA（AP003469.2和SPRY4-AS1）和两个靶基因（PPARGC1A和SLC2A1）（P<0.05），其中PPARGC1A高表达提示预后较好，AP003469.2、SPRY4-AS1和SLC2A1基因高表达则提示预后较差。基于4个基因构建的预后风险评分模型，第1、3和5年的AUC分别为0.730、0.699和0.679，提示模型具备良好的预测效能。在敲减AP003469.2后，SMMC-7721细胞中YWHAZ的表达无明显改变，且对细胞的增殖无影响。结论：基于生物信息学分析共筛选出4个关键基因，其中eRNA AP003469.2是HCC预后预测效能较高的标志物，其无促癌功能且对YWHAZ基因无调控作用。

Regorafenib regulates the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells through miR-122 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究瑞戈非尼（regorafenib，Rego）对人肝癌SMMC-7721细胞增殖、凋亡的影响及其可能的机制。方法：将SMMC-7721细胞分为对照组及Rego组，分别用0、10 μmol/L Rego处理48 h后，流式细胞术检测各组细胞凋亡率，qPCR检测细胞中miR-122的表达。采用脂质体转染的方法将合成的hsa-miR-122-5p模拟物转染SMMC-7721细胞构建miR-122过表达的overExp-miR-122细胞，并将细胞分为对照组、Rego组、overExp-NC组、overExp-NC+Rego组、overExp-miR-122组及overExp-miR-122+Rego组，采用MTT法检测细胞活性，流式细胞术检测细胞凋亡率、WB法检测细胞中Bcl2、cleaved caspase-3、RAS、RAF1、p-ERK1蛋白表达水平。结果：与对照组相比，Rego处理后细胞凋亡率显著升高（P<0.05），且miR-122表达量显著上升（P<0.01）；与overExp-NC组比较，overExp-miR-122组细胞增殖抑制率、凋亡率和cleaved caspase-3表达均显著升高（均P<0.01），RAS蛋白表达显著下降（P<0.05），Bcl2、RAF、p-ERK1蛋白表达均显著下降（均P<0.01）；与overExp-miR-122组相比，overExp-miR-122+Rego组细胞中各检测指标变化进一步显著增加（P<0.01）。结论：Rego可抑制SMMC-7721细胞增殖、促进凋亡，其作用可能与调控miR-122、凋亡相关因子的表达和抑制RAS/RAF/ERK信号通路有关。

Effect of melittin on programmed necrosis of human hepatocellular carcinoma cell line SMMC￣7721 / 解剖学报

Objective To investigate the lethal effect of melittin on human hepatocellular carcinoma cell line SMMC-7721 and its possible mechanism. Methods MTT assay was used to investigate the killing effect of different concentrations melittin on human hepatocellular carcinoma cells SMMC-7721. Meanwhile, the inhibitory effect of specific programmed necrosis inhibitors necrostain-1(Nec-1) on melittin killing SMMC-7721 cells was detected. Programmed cell necrosis observed by Hoechst 33342 and PI double staining. The necrotosis rate was detected by flow cytometry. Ultrastructural changes of cell was detected by transmission electron microscopy. The expression of receptor interacting protein 1(RIP1) was detected by Western blotting. Results Compared with the control group, the proliferation activity of SMMC-7721 cells was significantly decreased after treatment with different concentrations of melittin for 24 hours (P < 0. 05) . Cells stained in dark blue and red. Cell membrane integrity was destroyed, organelle swelling, organelle membrane was destroy, that demonstrates cell was necrosis. Westen blotting result showed an increased proportion of RIP1 expression in SMMC-7721 cells. Compared with the melittin group, cell proliferation activity was significantly increased, cell necrotosis rate was decreased, and intracellular RIP1 expression was down-regulated in the Nec-1 pretreatment group. Conclusion Melittin induces cell death in SMMC-7721 cells through the RIP1-mediated programmed necrosis pathway.

Over-expression of miR-144 inhibits invasion of liver cancer SMMC-7721 cells by suppressing TLR/MyD88 pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effects of over-expression of miR-144 on invasion of SMMC-7721 cells and Toll-like receptor (TLR)/myeloid differentiation factor 88 (MyD88) pathway in hepatocellular carcinoma cells.@*METHODS@#The expressions of miR-144 was examined in normal human hepatocyte line HL-7702 and hepatocarcinoma cell line SMMC-7721 using realtime quantitative PCR (qRT-PCR). SMMC-7721 cells were divided into blank group, miR-144 NC group and miR-144 mimics group, and the expressions of miR-144 in each group were detected with qRT-PCR. Cell count kit-8 (CCK8) was used to assess the survival of SMMC-7721 cells, and the cell invasion was evaluated using Transwell assay. The expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and TLR/MyD88 pathway-related proteins in the cells were detected with Western blotting; the effect of 40 μ mol/L MyD88 inhibitor on TLR/MyD88 pathway-related proteins was examined in SMMC-7721 cells.@*RESULTS@#Compared with normal human hepatocytes, SMMC-7721 cells expressed a significantly lower level of miR-144 ( < 0.05). CCK-8 assay showed that test showed that miR-144 over-expression significantly decreased the cell survival rate ( < 0.05), lowered the number of invasive cells, and decreased the expression of MMP-2 and MMP-9 in SMMC-7721 cells ( < 0.05). The expressions of Toll-like receptor 4 (TLR4), MyD88, phosphorylated nuclear factor-kappa B (pNF-κB) and NF-κB protein decreased significantly in miR-144 mimics group and TJ-M2010-2 group ( < 0.05) and were comparable between the two groups ( > 0.05).@*CONCLUSIONS@#Overexpression of miR-144 decreases SMMC-7721 cell survival and invasion by inhibiting TLR/MyD88 pathway.

miR-223-3p regulates proliferation and apoptosis of hepatocellular carcinoma SMMC-7721 cells by targeting RAC1 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of miR-223-3p on the proliferation and apoptosis of hepatocellular carcinoma (HCC) cells by regulating Ras-related C3 botulinum toxin substrate 1 (RAC1) and its possible mechanism. Methods: Thirty pairs of HCC and corresponding para-cancer tissues resected in Jilin Central Hospital from August 2016 to August 2018 were collected for this study; in addition, human HCC cell lines SMMC-7721, BEL-7402, HepG2 and human normal hepatocyte QSG-7701 were also collected. The expression level of miR-223-3p in HCC tissue and cell lines was detected by qPCR. miR-223-3p mimics, miR-223-3p inhibitor and siRAC1 were transfected into SMMC-7221 cells, respectively. CCK-8 assay, Colony formation assay and Annexin V-FITC/PI staining Flow cytometry were used to detect the proliferation, clone formation and apoptosis of SMMC-7721 cells, respectively. The relationship between miR-223-3p and RAC1 was confirmed by Dual luciferase reporter gene assay. The protein level of RAC1 in SMMC-7721 cells was detected by Western blotting. Results: The expression of miR-223-3p in HCC tissues was significantly lower than that in paracaner tissues (P<0.01), and had significant correlation with pathological characteristics, such as tumor size, TNM stage, EdmondsonSteiner grade (all P<0.05 or P<0.01). miR-223-3p expression in HCC cell lines was significantly lower than that in QSG-7701 cells with the lowest expression in SMMC-7721 cells. Dual luciferase reporter gene assay confirmed that RAC1 was a target gene of miR-223-3p, and miR-223-3p negatively regulated RAC1 expression. Over-expression of miR-223-3p significantly inhibited the proliferation and colony formation (P<0.05 or P<0.01) of SMMC-7721 cells and promoted cell apoptosis (P<0.01). Contrarily, knockdown of miR-223-3p reversed the inhibitory effect of miR-223-3p mimics on cells. Conclusion: miR-223-3p over-expression inhibits proliferation and colony formation and promotes apoptosis of HCC cells, the mechanism of which may be related with its targeted down-regulation of RAC1.

Reimaging biological barriers affecting distribution and extravasation of PEG/peptide- modified liposomes in xenograft SMMC7721 tumor

Liposomes, as one of the most successful nanotherapeutics, have a major impact on many biomedical areas. In this study, we performed laser scanning confocal microscope (LSCM) and immunohistochemistry (IHC) assays to investigate the intra-tumor transport and antitumor mechanism of GE11 peptide-conjugated active targeting liposomes (GE11-TLs) in SMMC7721 xenograft model. According to classification of individual cell types in high resolution images, biodistribution of macrophages, tumor cells, cells with high epidermal growth factor receptor (EGFR) expression and interstitial matrix in tumor microenvironment, in addition, their impacts on intra-tumor penetration of GE11-TLs were estimated. Type I collagen fibers and macrophage flooded in the whole SMMC7721 tumor xenografts. Tumor angiogenesis was of great heterogeneity from the periphery to the center region. However, the receptor-binding site barriers were supposed to be the leading cause of poor penetration of GE11-TLs. We anticipate these images can give a deep reconsideration for rational design of target nanoparticles for overcoming biological barriers to drug delivery.

Ampelopsin inhibited apoptosis and migration of SMMC-7721 cells through MAPKs signaling pathway / 药学学报

This research is aimed to investigate the effect of ampelopsin on apoptosis and migration of human hepatoma SMMC-7721 cells and explore the molecular mechanism. SMMC-7721 cells were pretreated with different doses of ampelopsin and cells proliferation was detected by CCK8 kit. Cell morphology was observed under an inverted microscope. Nuclear morphology was detected by DAPI staining. Apoptotic rate was detected by Annexin V-FITC/PI flow cytometry. Migration and invasion were detected by Transwell and scratch healing test. Western blotting was used to detect cleavage of poly ADP-ribose polymerase (PARP), expression of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), E-cadherin, and N-cadherin, and phosphorylation of ERK, P38 and JNK in MAPKs pathway. Our results showed that ampelopsin significantly inhibited proliferation and induced apoptosis of SMMC-7721 cells, with half inhibition dose (IC50) for 24 h was 38.98 μg·mL-1. With 50 μg·mL-1 ampelopsin treatment, typical apoptotic morphological changes occurred, such as cell detachment, shrinkage and nuclear condensation. Apoptotic rate increased from 15% to 55.1%, with PARP cleavage significantly increased. In addition, treatment of ampelopsin reduced scratch healing of cells and transmembrane cells number. The expression levels of MMP-2 and MMP-9 were decreased. Further analysis of EMT-related proteins showed that after ampelopsin treatment, E-cadherin was up-regulated and N-cadherin was down-regulated. During ampelopsin treatment, ERK reached its peak of activation after 1 h, while the maximum activation time of JNK was 12 h. Meanwhile, P38 was activated within 4 h, with the highest point at 2 h. But after 4 h, ampelopsin inhibited phosphorylation of P38. These results indicated that ampelopsin induced apoptosis and reduced migration through activating MAPKs pathway and reversing EMT process in SMMC-7721 cells. This work provides a mechanistic basis for utilizing ampelopsin for anti-hepatocarcinoma treatment.

The Research of Hyaluronic Acid Functionalized Mesoporous Silica Nanoparticles MCM-41 Loading Paclitaxel for SMMC-7721 Liver Cancer Cells / 中国药学杂志

OBJECTIVE: To establish a drug delivery system based on hyaluronic acid functionalized mesoporous silica nanoparticles MCM-41 loaded with paclitaxel (HA-MCM-41-PTX). The physical and chemical properties, in vitro drug release and the antitumor effect were investigated. METHODS: The morphological structure and particle size of MCM-41 were observed by TEM. The drug delivery system was characterized by PXRD and FTIR. The in vitro release experiments was carried out to investigate the dissolution rate of HA-MCM-41-PTX. The in vitro cells experiment was carried out to explore the mechanism of HA-MCM-41-PTX on cells. RESULTS: The drug loading capacity of HA-MCM-41-PTX was 28.75%. The in vitro drug release experiments showed that HA-MCM-41-PTX exhibited controlled release with a cumulative release of (86.19±5.11)% until 48 h. In vitro cell experiments showed that HA-MCM-41-PTX had excellent targeting effect due to the modification of hyaluronic acid, which was easier to be uptaken by cells and exhibited great antitumor effect. CONCLUSION: HA-MCM-41-PTX is an excellent drug delivery system with both controlled release and targeting antitumor effect.

Effect of Capsaicin on Proliferation and HMGB1 and IL-6 Expressions in Hepatoma SMMC-7721 Cells / 中国实验方剂学杂志

Objective: To investigate the effect of capsaicin on proliferation in human hepatoma SMMC-7721 cells and its possible molecular mechanism. Method: Capsaicin (50,100,150,200,250,300 μmol·L-1) groups and blank group were set up. The cell viability was detected by cell counting kit-8 (CCK-8) assay after SMMC-7721 cells were treated with capsaicin (50,100,150,200,250,300 μmol·L-1) for 24, 48, 72 h. The morphological changes were observed under an inverted microscope after SMMC-7721 cells were treated with capsaicin (150,200,250 μmol·L-1) for 24 h. The mRNA expression levels of high mobility group box 1 (HMGB1) and interleukin-6(IL-6) were measured by Real-time PCR after SMMC-7721 cells were treated with capsaicin (150,200,250 μmol·L-1) for 24 h. The levels of HMGB1 and IL-6 in cell culture supernatant were detected by enzyme-linked immunosorbent assay(ELISA) after SMMC-7721 cells were treated with capsaicin (150,200,250 μmol·L-1) for 24 h. Result: Compared with the blank group, there was no significant difference between 50 and 100 μmol·L-1 capsaicin groups treated for 24, 48, 72 h; after treated with the other concentrations of capsaicin (150, 200, 250, 300 μmo·L-1) at different time points, the proliferation inhibition rate was statistically significant (P-1) groups showed different degrees of morphological changes in SMMC-7721 cells, which became round and wrinkled, with a poor attachment and more exfoliation; compared with the blank group, the mRNA expressions of HMGB1 and IL-6 in SMMC-7721 cells of capsaicin (150, 200, 250 μmol·L-1) groups were significantly down-regulated (PPConclusion: Capsaicin inhibits cell proliferation of SMMC-7721 cells, and the possible mechanism may be related to the down-regulation of HMGB1 and IL-6 at the mRNA and protein levels.

Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics / 中国实验方剂学杂志

Objective: To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS). Method: CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0, 12.5, 25, 50, 100 μmol·L-1) on human liver cancer Huh7, SMMC-7721 cells for 24, 48, 72 h. GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells (Huh7, SMMC-7721) for 24 h. The data was preprocessed by Postrun Analysis 4.41 workstation. Partial least squares-discriminant analysis (PLS-DA) was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on MetaboAnalyst 3.0 software. Result: Compared with the normal group, after two kinds of liver cancer cells was treated by artesunate, a total of 39 identical metabolites in the cells have undergone significant changes, which were mainly related to five metabolic pathways,including biosynthesis of aminoacyl-transfer RNA (tRNA), metabolism of alanine, aspartic acid and glutamic acid, metabolism of glycine, serine and threonine, metabolism of arginine and proline, metabolism of glutathione. Conclusion: Artesunate (12.5-100 μmol·L-1) can inhibit the growth of liver cancer cells (Huh7, SMMC-7721), it mainly involves five metabolic pathways, which may be the pathway of artesunate against liver cancer.

Over-expression of PRRX1 induces apoptosis of hepatocellular carcinoma SMMC7721 cells through a p53-mediated mitochondrial apoptosis pathway / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of over-expression of paired related homoeobox 1 (PRRX1) on apoptosis of hepatocellular carcinoma SMMC7721 cells, and to explore its detailed mechanism. Methods： ：Lentivirus-mediated PRRX1 over-expression vector (pGMLV-PRRX1) and empty vector (Vector) were respectively infected into SMMC7721 cells, and the mRNA and protein expression levels of PRRX1 in infected cells were detected by qPCR and WB. The effect of PRRX1 over-expression on the cell proliferation and apoptosis of SMMC7721 cells were assessed by CCK-8 assay and Annexin-V FITC/PI double staining flow cytometry assay, respectively. The change of mitochondrial membrane potential of SMMC7721 cells was detected by mitochondrial membrane potential assay kit (JC-10 staining assay). The enzymatic activities of caspase-8 and caspase-9 in infected cells were detected by using caspase activity assay kit (spectrophotometric method). The protein expression levels of p53, Bcl-2, Bax, Fas, Cleaved-caspase-3 and Cty C expressed in mitochondria and cytosol were evaluated by WB. Results： ：PRRX1 over-expressed SMMC7721 cell line was successfully constructed, and the protein and mRNA expression levels of PRRX1 were significantly increased in lentivirus infected cells (all P< 0.01). Compared with control group and vector group, over-expression of PRRX1 significantly inhibited cell proliferation, weakened mitochondrial membrane potential, but increased the rate of apoptosis, elevated the shear level of caspase-3, promoted the release of Cyt C protein from mitochondrial into cytosol and increased the enzymatic activity of caspase-9 (all P<0.05 or P<0.01). In addition, over-expression of PRRX1 also promoted the protein expressions of p53 and Bax but inhibited the protein expression of Bcl-2 (all P< 0.05 or P<0.01); however,it had no significant effect on the expression of Fas protein and the enzymatic activity of caspase-8 (all P＞ 0.05). Conclusion： ：Over-expression of PRRX1 induces apoptosis in hepatocellular carcinoma SMMC7721 cells, which may be related to the activation of p53-mediated mitochondrial apoptotic pathway

Psoralen inhibits malignant proliferation and induces apoptosis through triggering endoplasmic reticulum stress in human SMMC7721 hepatoma cells

BACKGROUND: Psoralen is a coumarin-like and coumarin-related benzofuran glycoside, which is a commonly used traditional Chinese medicine to treat patients with kidney and spleen-yang deficiency symptom. Psoralen has been reported to show estrogen-like activity, antioxidant activity, osteoblastic proliferation accelerating activity, antitumor effects and antibacterial activity. However, the antitumor mechanism of psoralen is not fully understood. This study aimed to investigate the therapeutic efficacy of psoralen in human hepatoma cell line SMMC7721 and the mechanism of antitumor effects. RESULTS: Psoralen inhibited proliferation of SMMC7721 in a dose- and time-dependent manner, and promoted apoptosis. Further, psoralen activated the ER stress signal pathway, including the expansion of endoplasmic reticulum, increasing the mRNA levels of GRP78, DDIT3, ATF4, XBP1, GADD34 and the protein levels of GDF15, GRP78, IRE1α, XBP-1s in a time-dependent manner. Psoralen induces cell cycle arrest at G1 phase by enhancing CyclinD1 and reducing CyclinE1 expression. Moreover, TUDC couldn't inhibit the psoralen-induced ER stress in SMMC7721 cells. CONCLUSIONS: Psoralen can inhibit the proliferation of SMMC7721 cells and induce ER stress response to induce cell apoptosis, suggesting that psoralen may represent a novel therapeutic option for the prevention and treatment hepatocellular carcinoma.

Chrysin promotes SMMC-7721 cell apoptosis by regulating MAPKs signaling pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of chrysin in inducing apoptosis of human hepatic carcinoma cells and explore the possible mechanism.</p><p><b>METHODS</b>Human hepatic carcinoma SMMC-7721 cells treated with DMSO or chrysin at different concentrations (5-200 μg/mL) were examined for changes in the cell proliferation using CCK-8 assay. The morphological changes of SMMC-7721 cells were observed in response to treatment with 5, 10, or 20 μg/mL chrysin, and the changes in the cell nuclei were observed using DAPI nuclear staining. Annexin Ⅴ-FITC/PI flow cytometry was used to determine the cell apoptosis rate. The changes in the apoptosis-related proteins (PARP and caspase-3) and MAPKs signal pathway were detected with Western blotting.</p><p><b>RESULTS</b>Chrysin treatment obviously suppressed the proliferation of SMMC-7721 cells in a dose-dependent manner below the concentration of 60 μg/mL. Chrysin (20 μg/mL) also caused significantly increased cell apoptosis and significant cleavage of PARP and caspase-3. Chrysin significantly activated MAPKs signaling pathway in a time-and dose-dependent manner, with the peak activation level occurring at 15 min. Pretreatment of the cells with specific inhibitors of the MAPKs pathway obviously inhibited the effect of chrysin in inducing cell apoptosis.</p><p><b>CONCLUSIONS</b>Chrysin inhibits the proliferation and promotes apoptosis of SMMC-7721 cells by regulating the activation of MAPKs signaling.</p>

Effects of 3 Extracts of Acanthopanax sessiliflorus Fruits on the Proliferation and Apoptosis of Human Hepatocarcinoma Cells SMMC-7721 / 中国药房

OBJECTIVE:To study the effects of 3 extracts of Acanthopanax sessiliflorus fruits on the proliferation and apoptosis of human hepatocarcinoma cells SMMC-7721,and to provide reference for confirming the mechanism of anti-tumor effect. METHODS:MTT assay was adopted to investigate the effects of low-mass concentration,medium-mass concentration and high-mass concentration of ethanol extract(0.92,1.84,3.68 mg/mL),crude polysaccharide extract(0.06,0.12,0.24 mg/mL)and refined polysaccharide extract (0.04, 0.08, 0.16 mg/mL) from A. sessiliflorus fruits on the proliferation and apoptosis of SMMC-7721 cells after treated for 24,36,48 h,respectively. Flow cytometry was used to investigate the effects of 1.84 mg/mL ethanol extract,0.24 mg/mL crude polysaccharide extract and 0.16 mg/mL refined polysaccharide extract on cell cycle and cell apoptosis after treated for 24 h. The above tests were all negative control(only adding cells without drugs). RESULTS:Compared with negative control,3 extracts of A. sessiliflorus fruits could significantly inhibit the proliferation of SMMC-7721 cells (P<0.01),could significantly decrease the percentage of SMMC-7721 cells in G0/G1 and G2/M phase(P<0.01),could significantly increase the percentage of SMMC-7721 cells in S phase (P<0.01) and the apoptosis rate of SMMC-7721 cells (P<0.05);especially the effects of ethanol extract from A. sessiliflorus fruits were the most obvious. CONCLUSIONS:Three extracts of A.sessiliflorus fruits can inhibit the proliferation of human hepatocarcinoma SMMC-7721 cells,block SMMC-7721 cells in S phase and induce the apoptosis of SMMC-7721 cells.

Structure-activity relationship of annonaceous acetogenins against multidrug resistant human hepatic carcinoma cell line SMMC-7721/ADR / 中国中药杂志

The present research was launched to investigate the effects of 12 annonaceous acetogenins (ACGs) on human hepatic carcinoma cell line SMMC-7721/ADR, and to find out their structure-activity relationship. SMMC-7721/ADR cells were treated with 12 ACGs including annosquamin A(1)，annosquamin B(2)，annotemoyin-1(3)，uvariamicin Ⅱ(4)，annosquacin D(5)，annosquacin B(6)，isodesacetyluvaricin(7)，uvarigrandin A(8)，squamostatin D(9)，squamostatin E(10)，squamostatin A(11)，and 12,15-cis-squamostatin A(12) for 24 h, and the different expression of the target gene NDUFV2 were detected by quantitative real-time PCR. All the tested compounds made the expression of the target gene NDUFV2 decreased on human hepatic carcinoma cell line SMMC-7721/ADR, of which the bistetrahydrofuran ACGs showed the best activity,which the non-adjacent bistetrahydrofuran ACGs displayed the worst activity.The ACGs with the reducing number of carbons between γ-unsaturated lactone and the close tetrahydrofuran (THF) ring are more potent. For bistetrahydrofuran ACGs with the same nucleus skeleton,they would be more active as more hydroxyls on aliphatic chain, which for the non-adjacent bistetrahydrofuran ACGs with less hydroxyls on aliphatic chain that would be more active. ACGs with 3 hydroxyls on aliphatic chain would be more active. ACGs with threo configuration are more active than erythro configurotion, and the compounds with cis THF ring seem to be superior to those of trans THF ring. Furthermore, the ACGs with the reducing number of carbons between terminal methyl and the close tetrahydrofuran (THF) ring are more potent.

Effect of Marine Capsules on BALB/c Human Hepatocellular Carcinoma Cell Line SMMC-7721 in vitro / 中国药师

Objective:To observe the effect of marine capsules on BALB/c human hepatocellular carcinoma cell line SMMC-7721 in vitro. Methods:Human hepatoma SMMC-7721 cell line was cultured in vitro to establish a hepatoma tumor model, and was random-ly divided into the model control group, 5-Fu group (25 mg·kg-1), marine capsules high dose group (90 mg·kg-1), marine cap-sules low dose group (45 mg·kg-1 ) and the blank control group with 10 mice in each and 14-day continuous administration. Blood was taken from the eyeball and serum was isolated. The tumor strains were picked, the wet weight was obtained, the inhibition rate was calculated, and the pathological changes of tumor tissues were observed after HE staining. Results: Compared with that in the model control group, the serum IL-2 content significantly increased in the marine capsules high and the low dose groups ( P<0. 01, P<0. 05). The in vitro growth of tumor (P<0. 001, P<0. 05) was inhibited with the inhibition rate of 41. 56% and 27. 78% respectively for the high dose group and the low dose group, and the effects of the high dose group had no significant differences with those of 5-Fu group (P>0. 05). The nuclear staining of tumor cells was lighter and the number of tumor cells decreased in the two marine capsules groups. Conclusion:Marine capsules have certain inhibitory effects on BALB/c human hepatocellular carcinoma cell line SMMC-7721 in vitro.

Effect of myricetin on migation and invasion ofhepatoma SMMC-7721 cells / 中国药理学通报

Aim To investigate the effects of myricetin on the migration and invasion of hepatoma SMMC-7721 cells.Methods SMMC-7721 cells were treated with different concentrations of myricetin.Migration and invasion of SMMC-7721 cells were examined by Wound healing and Transwell, and the expression of E-cadherin and N-cadherin was detected by RT-qPCR and Western blot.Results Myricetin may inhibit the viability of SMMC-7721 cells via different concentration(10~40 μmol·L-1);furthermore, myricetin might also inhibit the migration and invasion of hepatoma SMMC-7721 cells.Meanwhile with the increasing of myricetin concentration, both filopodia and lamellipodia formation was reducd, and 7721 cells displayed in more integrity.And data from RT-qPCR and Western blot demonstrated that myricetin may up-regulate the expression of E-cadherin, simultaneously, down-regulate N-cadherin in SMMC-7721 cells.Conclusion Myricetin may influence the cell migration and invasion through up-regulating the expression of E-cadherin, simultaneously, down-regulating N-cadherin and activate cytoskeleton remolding of SMMC-7721.

Effects of volatile components in Oroxyli Semen on H22 bearing mice and its possible mechanisms / 中国药理学通报

Aim To study the inhibitory effect of volatile components in Oroxyli Semen on liver cancer and its possible mechanisms.Methods H22 bearing mouse model was used,the mice were divided into six groups:blank,model,positive (cytoxan,100 mg · kg-1),low-,mid-,and high-dose (17.5,35,and 70 mg · kg-1) volatile components groups,and then the mice were ig given once daily for consecutive 12 d.Then the tumor growth inhibitory rate,spleen and thymus indexes were calculated;the serum levels of IL-2,IL-6 were determined.HE staining was used to study of the apoptosis of the solid tumor.After treatment of SMMC-7721 cells with 0 ～ 1 g · L-1 of volatile components for 24,48 and 72 h,MTT assay was used to examine the proliferation.TUNEL method was applied to detect cell apoptosis,and RT-PCR method to detect Bax,Bcl-2,caspase-3 mRNA experssion.Results The inhibitory rate of volatile components high-dose on H22 bearing mice was 42.08％.The thymus index and the contents of serum IL-2 and IL-6 of H22 bearing mice were significantly higher than those in model group.Volatile components could significantly inhibit proliferation and induce apoptosis of SMMC-7721 cells,downregulate the expression of Bcl-2 mRNA,and up-regulate the expression of Bax,caspase-3 mRNA.Conclusions The volatile components in Oroxyli Semen have obvious anti-tumor activity in vitro and in vivo,and its mechanism may be related to enhancing immune system and promoting tumor cell apoptosis.