乳腺癌是全球最常见的癌症之一,严重威胁妇女健康.目前乳腺癌的治疗方法包括手术、化疗、放疗、内分泌治疗及分子靶向治疗等,其中利用铁死亡治疗乳腺癌是一种有前景的治疗策略.铁死亡是一种受调控的细胞死亡形式,其特征是铁依赖性的脂质过氧化物的积累.铁死亡与乳腺癌的发展密切相关,诱导铁死亡能抑制乳腺癌细胞增殖、侵袭和迁移,降低其对化疗药物的耐药性、增强其对放疗的敏感性.因此,诱导乳腺癌发生铁死亡是一种具有潜力和价值的治疗策略.

肝细胞癌是常见的恶性肿瘤,其发病机制尚未完全明确.Wnt信号通路与人体内多种病理生理过程相关,其中肝癌的发生、发展可能与经典的Wnt/β-catenin信号通路密切相关.Wnt/β-catein信号通路通过表达癌症相关基因、激活肝星状细胞、调控肝干细胞行为、促进肝癌细胞侵袭转移等方式调控肝癌的发生、发展.Wnt/β-catein信号通路在肝癌发生、发展中的作用有望为肝癌研究提供新的思路.

目的:探讨中药重楼(Paris polyphylla,PP)活性单体PP-11体外抑制人乳腺癌MDA-MB-231细胞增殖的作用及其机制.方法:采用不同浓度的PP-11作用于MDA-MB-231细胞,采用MTT法和集落形成实验检测细胞增殖情况;Hoechst 33258染色及Annexin V-FITC/PI双染流式细胞术检测细胞凋亡;JC-1染色检测线粒体膜电位改变;Western blot法检测细胞凋亡及自噬相关蛋白表达情况.再采用总caspase抑制剂Z-VAD-FMK、p38抑制剂SB203580和自噬抑制剂氯喹(chloroquine,CQ)进行阻断实验.结果:MTT测定结果显示,PP-11以剂量和时间依赖方式显著抑制MDA-MB-231细胞活力,其作用24、48和72 h后的IC50分别为5.64、4.58和3.06μmol/L.集落形成实验结果提示,PP-11显著抑制MDA-MB-231细胞集落形成.Hoechst 33258染色观察到PP-11组MDA-MB-231细胞出现典型的细胞凋亡形态.Annexin V-FITC/PI双染流式细胞术结果显示,随着PP-11浓度的增加,MDA-MB-231细胞凋亡率逐渐升高.JC-1染色结果显示,PP-11处理的MDA-MB-231细胞线粒体膜电位下降.Western blot检测结果显示,PP-11处理的MDA-MB-231细胞Bcl-2家族蛋白中抗凋亡蛋白Bcl-2和Bcl-xL的表达显著减少,促凋亡蛋白Bim和Bok表达增加,p-p38和p-p53蛋白水平升高,p-ERK蛋白水平降低,cleaved caspase-9、cleaved cas?pase-3及cleaved PARP的蛋白水平显著升高;PP-11降低MDA-MB-231细胞p-STAT3蛋白水平及其下游蛋白c-Myc、cyclin D和Mcl-1的表达.总caspase抑制剂Z-VAD-FMK和p38 MAPK抑制剂SB203580均可减弱PP-11诱导的细胞凋亡.随着PP-11作用浓度的增加,细胞自噬相关蛋白LC3-Ⅱ表达增加,p62/SQSTM1表达下降;采用PP-11联合自噬阻断剂CQ,可逆转PP-11诱导的cleaved PARP表达,并提高细胞活力(P<0.05).结论:重楼单体PP-11可显著抑制人乳腺癌MDA-MB-231细胞增殖.PP-11通过激活p38 MAPK信号通路、抑制ERK和JAK-Stat3信号通路诱导MDA-MB-231细胞发生线粒体相关的凋亡,并促进细胞自噬.

目的 探讨葡萄膜黑色素原位瘤与转移瘤差异表达基因及其作用;探索乳腺癌1号基因相关蛋白1(BAP1)缺失的葡萄膜黑色素瘤(UM)差异表达基因以及预测BAP1靶标,探讨BAP1缺失对UM转移作用及其机制.方法 生信分析筛选UM转移差异表达基因及其功能,对公共数据库GEO中的数据集GSE27-831,GSE48863以及GSE39171中芯片数据以2为底数进行对数转换以及标准化(P＜0.05,|logFC|＞1)等预处理.为了进一步捕捉术语之间的关系,选择丰富的术语的子集,并将其作为网络图绘制,其中相似性＞0.3的项由边缘连接.我们从20个簇中的每个簇中选择具有最佳P值的项,约束每个集群不超过15个术语,总共不超过250个术语.该网络用CytoSCAPE可视化插件,其中每个节点表示1个丰富的术语,并用其簇的名称着色.对于每个给定的基因列表,利用以下数据库进行蛋白质-蛋白质相互作用富集分析:BioGrid、InWeb_IM、OmniPath.所得到的网络至少包含与另一个列表成员形成交互的蛋白质子集.如果网络中包含3 ～500个蛋白质,则进一步应用分子复合检测(MCODE)算法来识别紧密连接的网络组件.路径和过程富集分析被独立地应用于每个MCODE组件,并且保留3个最佳评分(按P值,P＜0.05,FDR＜0.001生物途径视为有价值)术语作为对应组件的功能描述.结果 我们运用生物信息学技术对转移性和非转移性UM组织样本和BAP1沉默和正常表达UM细胞样本基于微阵列对基因组数据进行基因本体富集(GO)分析和京都基因和基因组百科全书(KEGG)分析,结果显示差异基因主要富集在补体凝血级联途径.结论 BAP1在转移性UM的异常低表达,UM中BAP1突变可能通过影响肿瘤血管生成促进肿瘤浸润转移.

Objective: To investigate pp-10,a monomer purified from Paris Polyphylla,induced apoptosis of human gastric carcinoma BGC-823 cells and the possible mechanism behind this effect. Methods: The effects of pp-10 on the proliferation of BGC-823 Cells were evaluated by the MTT assays and colony formation assays. The cellular morphology changes of BGC-823 cells were observed by fluorescence microscopy after Hoechst 33342 staining. Cell apoptosis was detected by Annexin V-FITC / Propidium iodide( PI) double staining using flow cytometry. The expression of apoptotic and autophagy associated proteins as LC3,P62,Caspase-9,Caspase-3,the Caspase substrate poly( ADP-ribose) polymerase( PARP),Bcl-2 proteins family such as Bax,Bcl-2,PI3 k /Akt signaling pathway numbers such as Akt,p-Akt,m TOR,p-m TOR,P70s6 k and p-P70s6 k were detected by Western blotting. Results: Paris Polyphylla monomer pp-10 inhibited the proliferation of BGC-823 cells significantly in dose- and time- dependent manner( P 0. 05). Cell clone formation inhibition assay demonstrated that cell clones decreased with the increase of drug concentrations. Apoptotic morphology such as cell shrinkage,nuclear condensation,nuclear fragmentation,chromatin condensation were also observed by staining with Hoechst33342 under fluorescence microscope. Flow cytometry analysis showed an increase of the percentage of apoptotic cells in a dose-dependent manner treated with pp-10 for 24 h. Active bands of mitochondria associated apoptotic signaling pathway proteins such as Caspase-3,Caspase-9 and PARP could be seen with western blotting analysis after BGC-823 cells were incubated with different concentrations of pp-10 for24 h. In addition,pp-10 upregulated the expression of Bax,a pro-apoptotic protein,down-regulated the expression of Bcl-2 protein,an anti-apoptotic protein,up-regulated the expression of LC3,down-regulated the expression of P62 protein,and also down-regulated PI3 K / Akt signal pathway as decreasing the phosphorylation of p-m Tor,p-p70S6 K,downstream of Akt. Conclusions: pp-10 inhibits cell proliferation and induces the apoptosis and autophagy of human Gastric carcinoma BGC-823 cells by activation of Bax,down-regulation of Bcl-2 and reduction of PI3 K / Akt signaling pathway.

Aim:To investigate the inhibitory effect of Harmine hydrochloride on human pancreatic cancer cell line AsPC-1 and the possible mechanism.Methods: MTT and cell colony formation inhibitory assay were used to investigate the inhibition effect of Harmine hydrochloride on human pancreatic cancer cell line AsPC-1.Changes in apoptotic cell percentage were calculated by flow cytometry.Western blotting assay was used to determine cell apoptosis and autophagy.Results: MTT showed that Harmine hydrochloride could inhibited human pancreatic cancer AsPC-1cells in a dose-dependent manner.IC50 of Harmine hydrochloride on AsPC-1cells at 24 h,48 h and 72 h was 116.5 μmol·L-1,66 μmol·L-1,22.3 μmol·L-1 respectively;Cell clone formation inhibition assay demonstrated that cell clones were decreased with increase of the concentration of Harmine hydrochloride.Annexin V-FITC/PI staining also showed a dose-dependent effect on late apoptotic and necrotic cells.Western blotting assay showed that the apoptosis and autophagy landmark protein Caspase-3,PARP,LC3-II and P62 protein expression levels changed gradually with increasing of Harmine hydrochloride.Conclusion:Harmine hydrochloride inhibits the cell survival of human pancreatic cancer cell AsPC-1 in a dose-dependent manner.Harmine hydrochloride inhibits cell growth by inducing apoptotic and autophagy.

AIM: To investigate the effects of artesunate on proliferation and apoptosis in human hepatocelluar carcinoma cell line HepG2 and to study the sensitizing effect of artesunate on HepG2 cells to chemotherapeutic drugs.METHODS: The proliferation of HepG2 cells was determined by the assay of cell counting kit-8(CCK-8) and the colony formation test.The morphology of HepG2 cells with Hoechst 33258 staining was observed under fluorescent microscope.Annexin V/propidium iodide(PI) was used to analyze the apoptosis and the cell cycle.The sensitizing effects of artesunate on HepG2 cells to chemotherapeutic drugs [5-fluorouracil(5-FU),carboplatin and epirubicin] were determined by CCK-8 assay.RESULTS: When treated with artesunate for 48 h,the proliferation of HepG2 cells was significantly inhibited in a dose-dependent manner.The IC50 was 19.2 μmol/L.Compared with the control cells,the colony formation of HepG2 cells treated with artesunate for 7 days was significantly inhibited.The nuclear fragmentation,karyopyknosis,chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with artesunate were observed.The cells in G2 phase increased obviously,and the percentages of hypodiploid cells and early apoptotic rates were significantly higher in artesunate treatment groups than those in control group.The IC50 of 5-FU,carboplatin and epirubicin combined with artesunate was 3.33,2.02 and 1.71 times sensitized as compared with control group,respectively.CONCLUSION: Artesunate effectively inhibits proliferation and induces apoptosis of HepG2 cells.Aresunate also sensitizes HepG2 cells to chemotherapeutic drugs.

BACKGROUND: Harmine has antitumor and antinociceptive effects, and inhibits human DNA topoisomerase. However, no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma. This study aimed to investigate the effects of harmine on proliferation and apoptosis, and the underlying mechanisms in the human hepatocellular carcinoma cell line HepG2.METHODS: The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation test. The morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining. Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell cycle. Western blotting was used to assess expression of the apoptosis-regulated genes Bcl-2, Bax, Bcl-xl, Mcl-1, caspase-3, and caspase-9. Mitochondrial transmembrane potential (Psi(m)) was determined using JC-1.RESULTS: Harmine inhibited the proliferation of HepG2 cells in a dose-dependent manner. Hoechst 33258 staining revealed nuclear fragmentation and chromosomal condensation, cell shrinkage, and attachment loss in HepG2 cells treated with harmine. The percentage of the sub/G1 fraction was increased in a concentration-dependent manner, indicating apoptotic cell death. PI staining showed that harmine changed the cell cycle distribution, by decreasing the proportion of cells in G0/G1 and increasing the proportion in S and G2/M. Harmine induced apoptosis in a concentration-dependent manner, with rates of 20.0%, 32.7% and 64.9%, respectively. JC-1 revealed a decrease in Psi(m). Apoptosis of HepG2 cells was associated with caspase-3 and caspase-9 activation, down-regulation of Bcl-2, Mcl-1, and Bcl-xl, and no change in Bax.CONCLUSIONS: Harmine had an anti-proliferative effect in HepG2 cells by inducing apoptosis. Mitochondrial signal pathways were involved in the apoptosis. The cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma.