柴胡对人食管癌细胞株Eca-109的抑制作用

目的:探讨中药柴胡对人食管癌细胞株Eca-109的抑制作用。方法:体外培养人食管癌细胞株(Eca-109),分别给以不同浓度的柴胡溶液对体外培养的Eca-109细胞株进行干预。以MTT比色法测定柴胡溶液对Eca-109细胞株的抑制增殖的作用,计算其细胞增殖存活率。结果:MTT实验显示柴胡溶液可以明显抑制Eca-109细胞株的增殖。用药后Eca-109细胞增殖存活率下降幅度与剂量呈正相。结论:中药柴胡在体外具有抗肿瘤作用,其机制是抑制肿瘤细胞增殖,及抑制细胞的分化而达到抗肿瘤的目的。     

CDCA3在食管癌中的表达及其对食管癌细胞增殖与凋亡的影响

目的研究细胞周期相关因子(CDCA3)在食管癌组织和细胞中的表达及其对食管癌细胞增殖与凋亡的影响。方法实时荧光定量PCR(qPCR)检测50例食管癌组织及其配对的癌旁组织中CDCA3 mRNA的表达水平,同时检测CDCA3 mRNA在不同食管癌细胞系(Eca109、Kyse-170、TE1)和人食管鳞状上皮细胞系Het-1A中的表达水平;采用si-CDCA3-1、si-CDCA3-2转染Eca109和TE1细胞,另转染si-NC作为阴性对照,MTS法检测细胞增殖能力,流式细胞仪检测细胞周期和凋亡变化;Western blot检测CDCA3、CyclinD1和cleaved caspase-3蛋白的表达变化。结果食管癌组织中CDCA3 mRNA的表达高于癌旁组织(P<0.05),Eca109、Kyse-170和TE1细胞中CDCA3 mRNA表达高于人食管鳞状上皮细胞(P<0.05)。沉默CDCA3表达后,Eca109细胞和TE1细胞的增殖率降低,G0/G1期细胞比例增加(P<0.05)。沉默CDCA3表达后,Eca109细胞和TE1细胞的凋亡率增加(P<0.05)。沉默CDCA3表达可降低CyclinD1蛋白表达和增加cleaved caspase-3蛋白表达(P<0.05)。结论CDCA3 mRNA在食管癌组织和细胞系中均高表达,干扰CDCA3的表达可能通过降低CyclinD1蛋白表达抑制细胞增殖,增加cleaved caspase-3蛋白表达促进细胞凋亡,提示CDCA3可能为食管癌的潜在治疗靶标。     

沉默Twist基因对食管癌细胞生长迁徙和侵袭的影响

目的：探讨Twist基因作为食管癌治疗靶点的可能性。方法采用westernblot方法检测Twist基因在食管癌组织和细胞（TE-1和Eca-109）中的表达;采用携带Twist-shRNA的慢病毒（Lenti-Twist-shRNA）沉默Twist基因表达;采用CCK-8方法检测Twist基因沉默后食管癌细胞的生长状况;采用动物模型检测Twist基因沉默后食管癌荷瘤的生长状况;采用流式细胞仪检测Twist基因沉默后食管癌细胞的凋亡变化。结果 Twist基因在食管癌组织和细胞中表达显著升高;Lenti-Twist-shRNA处理后（即沉默Twist基因后）,食管癌细胞的生长受到显著抑制;Lenti-Twist-shRNA处理后,食管癌荷瘤生长速度显著降低;Lenti-Twist-shRNA处理后,食管癌细胞发生显著凋亡。结论 Twist基因参与调控食管癌细胞的生长和凋亡,可以作为食管癌治疗的潜在靶点。     

MiR-101在乳腺癌中的表达及其意义

目的:探讨miR-101在乳腺癌组织中的表达变化。方法:收集乳腺癌组织及对应的癌旁组织,以qRT-PCR方法测定miR-101在乳腺癌组织中的表达变化。在乳腺癌细胞中转染miR-101 mimics、mimics control,以qRT-PCR方法检测上调效果,MTT方法测定细胞增殖变化,流式细胞术测定细胞凋亡变化,Transwell小室测定细胞迁移和侵袭数目变化,蛋白质印迹法测定细胞中cleaved caspase-3和MMP-2蛋白表达变化。结果:miR-101在乳腺癌组织中的表达水平明显低于癌旁组织(P<0.05)。与miR-NC比较,miR-101细胞中miR-101表达水平升高,细胞增殖能力降低,细胞凋亡率升高,细胞侵袭和迁移数目下降,细胞内的cleaved caspase-3蛋白水平表达升高,MMP-2蛋白表达水平减少(P<0.05)。结论:MiR-101在乳腺癌组织中表达下调,上调miR-101抑制乳腺癌细胞增殖、侵袭和迁移并诱导细胞凋亡。     

EGF对食管鳞癌细胞HIF-1α基因表达及功能的影响

目的 研究表皮生长因子（EGF）对人食管鳞癌细胞株HIF-1α基因表达及功能的影响及其对血管生成拟态相关基因表达的影响.方法 将人食管鳞癌细胞株Eca-109、TE13分为试验组（加入含EGF 100 μg/L的无血清DMEM培养液）和对照组（加入无血清DMEM培养液）,分别于常氧及缺氧下培养.应用蛋白质印迹法（Western blot）检测细胞HIF-1α蛋白与血管内皮钙黏附素（sVE-cadherin）、层黏连蛋白（Laminin5γ2）、酪氨酸蛋白激酶受体（EphA2）和基质金属蛋白酶-2（MMP-2）基因蛋白的表达,应用逆转录聚合酶链反应法（RT-PCR）检测HIF-1α与sVE-cadherin、EphA2和MMP-2 RNA的表达,基质胶（Matrigel）三维培养显微镜下观察并计数两组细胞的管状结构.结果 常氧和缺氧情况下,与对照组比较,试验组Eca-109、TE13细胞HIF-1α蛋白与sVE-cadherin、Laminin5γ2、EphA2基因蛋白的表达及sVE-cadherin、EphA2 RNA水平的表达均显著增加,差异均有统计学意义（P＜0.05）;而对MMP-2基因蛋白的表达及RNA水平的表达无明显影响,差异无统计学意义（P＞0.05）.Eca-109、TE13细胞均可形成典型的管网状结构,试验组在EGF作用下数目增加,与对照组比较差异有统计学意义（P＜0.05）.结论 在常氧和缺氧情况下EGF均可促进Eca-109、TE13细胞HIF-1α蛋白与sVE-cadherin、EphA2等基因蛋白及RNA的表达,均可形成典型的管网状结构,且EGF能有效促进肿瘤细胞体外血管生成拟态形成.     

人参皂苷Rg3对缺氧诱导的Eca-109和786-0细胞血管内皮生长因子和核因子κB表达的影响

目的 探讨人参皂苷Rg3对人食管癌细胞株Eca-109和人肾癌细胞株786-0细胞缺氧诱导的血管内皮生长因子(VEGF)和核因子KB(NF-kB)表达的影响.方法 在常氧和缺氧环境下,采用噻唑蓝(MTT)法检测Rg3对Eca-109和786-0细胞增殖的影响,实时荧光定量聚合酶链反应(PCR)检测Rg3对VEGF mRNA表达的影响,酶联免疫吸附测定(ELISA)法检测Rg3和NF-kB抑制剂对VEGF蛋白表达的影响,蛋白质印迹(Western)检测Rg3对P65蛋白表达的影响.结果 在常氧或缺氧环境中,MTT显示Rg3都能显著抑制Eca-109和786-0细胞增殖,实时荧光定量PCR显示Rg3能显著抑制Eca-109和786-0细胞VEGF mRNA表达,ELISA显示Rg3和NF-kB抑制剂都能显著抑制VEGF蛋白的分泌,Western显示Rg3抑制P65蛋白表达.结论 Rg3可以抑制缺氧和常氧状态下的VEGF表达,其机制与抑制P65蛋白表达密切相关.     

端粒酶反义寡核苷酸对食管癌细胞Eca-109端粒酶活性及细胞凋亡的影响

目的探讨人端粒酶反义寡核苷酸(PS-ASODN)对食管癌Eca-109端粒酶活性及细胞凋亡的影响。方法1-5uM的PS-ASODN、5uM的N-ASODN作用于Eca-109细胞后,在倒置显微镜下连续观察Eca-109细胞形态学改变;采用半定量TRAP-银染法检测端粒酶活性;用流式细胞仪检测细胞凋亡率及细胞周期变化。结果PS-ASODN对Eca-109细胞具有生长抑制作用,PS-ASODN作用后,细胞的生长速度缓慢,细胞体积缩小,部分细胞变形漂起。随着药物浓度增加,PS-ASODN对Eca-109细胞端粒酶活性的抑制作用逐渐增强,呈浓度依赖性和序列特异性。流式细胞仪检测到凋亡峰并受阻于G0/G1期,而对照组寡核苷酸无上述作用。结论PS-ASODN不但抑制食管癌的增殖,降低端粒酶的活性,而且具有促凋亡的作用,对食管癌的临床治疗具有重要的价值。     

Investigation of the anti-glioma activity of Oviductus ranae protein hydrolysate

Oviductus Ranae is the dry oviducts of Rana temporaria chensinensis, and it has been reported to have a range of biological activities. This study aimed to investigate the effects of Oviductus Ranae protein hydrolysate (ORPH) on human glioma C₆ cell proliferation and apoptosis in vitro and in vivo. Following in vitro treatment, cell viability and colony formation assays showed that ORPH inhibited C₆ cell proliferation. In addition, the results of western blotting also demonstrated that ORPH effectively regulated the expression of the apoptosis related proteins, cleaved caspase-3, Bax and Bcl-2, DNA staining and flow cytometry analysis demonstrated that ORPH significantly promoted apoptosis in this cell line, a finding that was confirmed in vivo using terminal deoxynucleotidyl transferase dUTP nick end labeling. Further investigation demonstrated that ORPH increased apoptosis by modulating the release of inflammatory cytokines and the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway; this was demonstrated using a PI3K/AKT inhibitor (NVP-BEZ235). In summary, the present study suggested that ORPH promoted apoptosis and inhibited glioma cell proliferation by influencing the PI3 K/AKT signaling pathway.     

Retracted Article: SNHG3 promotes proliferation and invasion by regulating the miR-101/ZEB1 axis in breast cancer

Background: Dysregulated lncRNA expression contributes to the pathogenesis of human tumors via the lncRNAs functioning as oncogenes or tumor suppressors. Small nucleolar RNA host gene 3 (SNHG3) was demonstrated to be upregulated in breast cancer cells. However, the detailed roles and molecular mechanism of SNHG3 in breast cancer are largely unknown. Methods: The expression of SNHG3, miR-101, and zinc finger E-box-binding protein 1 (ZEB1) in breast cancer tissues and cells was detected using qRT-PCR. The effects of SNHG3 on cell proliferation and invasion were evaluated using MTT, EdU, and cell invasion assays. The protein levels of Ki-67, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase MMP-2, and MMP-9 were analyzed using western blot analysis. A luciferase reporter assay and RNA immunoprecipitation (RIP) were performed to explore the interaction between SNHG3, ZEB1 and miR-101. A subcellular fractionation assay was used to detect the subcellular location of SNHG3. Xenograft tumor experiments were conducted to verify the role and mechanism of SNHG3 in breast cancer in vivo. Results: SNHG3 expression was upregulated in breast cancer tissues and correlated with poor prognosis. SNHG3 knockdown suppressed breast cancer cell proliferation and invasion, which was further demonstrated by high levels of proliferation marker proteins Ki-67/PCNA and metastasis-related proteins MMP-2/MMP-9. Additionally, SNHG3 was located in the cytoplasm of breast cancer cells. SNHG3 functioned as a molecular sponge for miR-101 in breast cancer cells. miR-101 was downregulated in breast cancer tissues and negatively correlated with SNHG3 expression. Moreover, ZEB1, a target of miR-101, was positively regulated by SNHG3 in breast cancer cells. ZEB1 mRNA expression was upregulated in breast cancer tissues and positively correlated with SNHG3 expression. Mechanistically, SNHG3 knockdown suppressed cell proliferation and invasion by upregulation of miR-101 and downregulation of ZEB1 expression in breast cancer cells in vitro and in vivo. Conclusion: SNHG3 promoted proliferation and invasion by regulating the miR-101/ZEB1 axis in breast cancer.

In the present study, we investigated the expression and functional roles of SNHG3 in breast cancer cells, as well as the underlying mechanism of SNHG3 involved in the progression of breast cancer in vitro and in vivo.     

5-氮杂-2-脱氧胞苷通过上调MGMT表达增加5-氟尿嘧啶联合紫杉醇的化疗敏感性

目的研究5-氮杂-2-脱氧胞苷(5-Aza-d C)与传统化疗药物5-氟尿嘧啶联合紫杉醇(5-FU+PTX)对人胃癌细胞生长的影响,探讨5-Aza-d C抗肿瘤的机制。方法培养人胃癌细胞株MKN-45,分为对照组、5-Aza-d C组、5-FU+PTX组和5-Aza-d C+5-FU+PTX组,利用CCK-8细胞增殖检测试剂盒、流式细胞术检测各组细胞的增殖和凋亡情况,利用RT-PCR和Western blot方法分别检测各组胃癌细胞的MGMT基因m RNA与蛋白表达,及凋亡蛋白cleaved caspase-3的表达。结果 5-Aza-d C及5-FU联合紫杉醇均可抑制MKN-45细胞的生长,48 h细胞存活率分别为(71.60±5.77,57.00±6.09)%,与5-FU+PTX组比较,三药联合使用后效果更明显(42.85±2.29)%,P=0.02;5-Aza-d C及5-FU联合紫杉醇组细胞凋亡率[(7.33±0.34)%,(13.06±1.67)%]及凋亡蛋白cleaved caspase-3水平[(43.00±4.00)%,(45.67±4.50)%]均增加,三药联合用药组凋亡率[(17.31±1.05)%]和cleaved caspase-3水平[(174.67±6.50)%]的表达更高。与对照组抑癌基因MGMT m RNA和蛋白表达量[(23.10±2.15)%、(10.47±1.39)%]比较,5-Aza-d C单独处理的MKN-45细胞MGMT基因m RNA(56±5.57)%及蛋白表达水平(20.33±5.50)%均升高,三药联合用药组MGMT升高程度更加明显[(75.67±6.50)%,(174.67±6.50)%],P<0.001,而5-FU联合紫杉醇组未发现明显变化。结论5-Aza-d C可以有效抑制胃癌细胞的生长,它与化疗药物5-FU和紫杉醇联合应用时抗肿瘤效应更加明显。     

Polyporus umbellatus inhibited tumor cell proliferation and promoted tumor cell apoptosis by down-regulating AKT in breast cancer

Breast cancer (BC) is the foremost cause of cancer-related mortality in women worldwide. Polyporus umbellatus is a polysaccharide preparation of the Chinese traditional herb medicine, which has been explored as an inhibitory compounds in suppressing many cancers. And AKT has been known as an essential signaling pathway to regulate cell proliferation and apoptosis via Mdm2/p53 and Caspase-3 signaling pathways respectively. In our study, western blot, RT-PCR, immunochemical assay, immunofluorescence as well as flow cytometry were performed in vitro or in vivo to determine the effects of Polyporus umbellatus on the progression of human laryngeal cancer. First, the breast cancer cell growth, invasion and migration were inhibited, as well as the tumor volume in nude mice was down-regulated for Polyporus umbellatus use. Additionally, our data also showed that Polyporus umbellatus suppressed breast cancer cells proliferation, which was linked with the down-regulation of AKT activation by Polyporus umbellatus treatment. Mdm was inactivated while p53 was stimulated for Polyporus umbellatus administration, displaying inhibitory role in tumor growth. Furthermore, Polyporus umbellatus could up-regulate breast cancer cells in G0/G1 phase during cell cycle, and at the same time reducing cells in S phase. Also, flow cytometry and western blot assays suggested that apoptosis was induced by the administration of Polyporus umbellatus, which enhanced Caspase-3 expressions by AKT-regulated anti-apoptotic and pro-apoptotic signals. In conclusion, our data indicated that Polyporus umbellatus had a potential role in controlling human breast cancer through inhibiting tumor cell proliferation, inducing apoptosis regulated by AKT, which might provide a therapeutic strategy for breast cancer suppression in the future.     

人食管癌间充质干细胞对食管癌细胞株Eca-109侵袭性的影响

目的探讨人食管癌间充质干细胞(hEC-MSCs)对食管癌细胞株Eca-109侵袭性的影响。方法在体外将间质干细胞与食管癌细胞株ECA-109非接触共培养,使用RT-PCR和Western blotting的方法检测间质干细胞对ECA-109细胞株中基质金属蛋白酶-9(MMP-9)和抑癌基因E-cadherin表达的影响。结果间质干细胞可明显上调ECA-109细胞株中的MMP-9表达,显著下调ECA-109细胞株中E-cadherin的表达。结论食管癌间充质干细胞可能参与调节食管癌细胞的侵袭与转移。     

CXCL12/CXCR4轴通过诱导miR-125b促进肝癌细胞肿瘤干性和5-氟尿嘧啶抵抗的机制

目的探究趋化因子CXCL12/趋化因子受体CXCR4轴通过诱导miR-125b促进肝癌细胞肿瘤干性和5-氟尿嘧啶(5-FU)抵抗的机制。方法选择人肝细胞癌细胞系Huh7,分为7组:对照组(正常培养的肝细胞癌系),CXCR4转染组(CXCR4模拟转染超表达),CXCR4沉默组(CXCR4低表达或者不表达),miR-125b转染组(miR-125b超表达),CXCL12+125b抑制剂组(CXCL12超表达的+抑制miR-125b的表达),5-FU处理组(10μg/L的5-FU处理细胞),5-FU+miR-125b转染组(10μg/L的5-FU处理+miR-125b超表达的细胞)。聚合酶链式反应分析miR-125b mRNA表达;蛋白质印迹法检测E-钙粘蛋白、波形蛋白、CXCR4蛋白、Caspase-3、Bcl-2和Bax的蛋白表达;Transwell分析各组细胞中的细胞迁移、侵袭测定;MTT检测细胞存活力;细胞计数试剂盒8(CCK-8)评估细胞增殖。结果与对照组相比,CXCR4转染组miR-125b mRNA、波形蛋白、CXCR4的蛋白表达显著升高,E黏着蛋白表达显著降低(P<0.05);与CXCR4转染组相比,CXCR4沉默组miR-125b mRNA、波形蛋白、CXCR4的蛋白表达显著降低,E黏着蛋白表达显著升高(P<0.05)。与对照组相比,miR-125b转染组细胞迁移、侵袭数量、细胞活力均显著增加,细胞凋亡率显著降低(P<0.05);与miR-125b转染组相比,CXCL12+125b抑制剂组,细胞迁移、侵袭数量、细胞活均显著减少,细胞凋亡率显著升高(P<0.05)。与对照组相比,5-FU处理组细胞增殖率、Bcl-2表达显著降低,caspase-3、Bax的蛋白表达显著升高(P<0.05),与5-FU处理组相比,5-FU+miR-125b转染组细胞增殖率、Bcl-2表达显著升高,caspase-3、Bax的蛋白表达显著降低(P<0.05)。结论MiR-125b通过激活CXCL12/CXCR4轴而被上调,miR-125b增强CXCR4的表达,这在触发肿瘤侵袭和进展中形成了正反馈回路。这些结果为miR-125b在肝癌进程和化学抗性的发展中的作用提供了新的线索,为肝癌的治疗提供了潜在的治疗靶点。     

The novel anthraquinone derivative IMP1338 induces death of human cancer cells by p53-independent S and G2/M cell cycle arrest

To identify novel small molecules that induce selective cancer cell death, we screened a chemical library containing 1040 compounds in HT29 colon cancer and CCD18-Co normal colon cells, using a phenotypic cell-based viability assay system with the Cell Counting Kit-8 (CCK-8). We discovered a novel anthraquinone derivative, N-(4-[{(9,10-dioxo-9,10-dihydro-1-anthracenyl)sulfonyl}amino]phenyl)-N-methylacetamide (IMP1338), which was cytotoxic against the human colon cancer cells tested. The MTT cell viability assay showed that treatment with IMP1338 selectively inhibited HCT116, HCT116 p53^−/−, HT29, and A549 cancer cell proliferation compared to that of Beas2B normal epithelial cells. To elucidate the cellular mechanism underlying the cytotoxicity of IMP1338, we examined the effect of IMP1338 on the cell cycle distribution and death of cancer cells. IMP1338 treatment significantly arrested the cell cycle at S and G2/M phases by DNA damage and led to apoptotic cell death, which was determined using FACS analysis with Annexin V/PI double staining. Furthermore, IMP1338 increased caspase-3 cleavage in wild-type p53, p53 knockout HCT116, and HT29 cells as determined using immunoblotting. In addition, IMP1338 markedly induced the phosphorylation of histone H2AX and Chk1 in both cell lines while the combination of 5-fluorouracil (5-FU) and radiation inhibited the viability of HCT116, HCT116 p53^−/−, and HT29 cells compared to 5-FU or radiation alone. Our findings indicated that IMP1338 induced p53-independent cell death through S and G2/M phase arrest as well as DNA damage. These results provide a basis for future investigations assessing the promising anticancer properties of IMP1338.     

Soluble Vascular Endothelial Growth Factor Decoy Receptor FP3 Exerts Potent Antiangiogenic Effects

The binding of vascular endothelial growth factor (VEGF) to its receptors stimulates tumor growth; therefore, modulation of VEGF would be a viable approach for antiangiogenic therapy. We constructed a series of soluble decoy receptors containing different VEGF receptor 1 (FLT1) and VEGF receptor 2 (KDR) extracellular domains fused with the Fc region of human immunoglobulin (Ig) and evaluated their antiangiogenic effects and antitumor effects. Results of in vitro binding and cell proliferation assays revealed that decoy receptor FP3 had the highest affinity to VEGF-A and -B. Compared with bevacizumab, FP3 more effectively inhibited human umbilical vein endothelial cell (HUVEC) migration and vessel sprouting from rat aortic rings. FP3 significantly reduced phosphorylation of AKT and ERK1/2, critical proteins in the VEGF-mediated survival pathway in endothelial cells. Moreover, FP3 inhibited tumor growth in human hepatocellular carcinoma (HepG2), breast cancer (MCF-7), and colorectal cancer (LoVo) tumor models, and reduced microvessel density in tumor tissues. The FP3-mediated inhibition of tumor growth was significantly higher than that of bevacizumab at the same dose. FP3 also demonstrated synergistic antitumor effects when combined with 5-fluorouracil (5-FU). Taken together, FP3 shows a high affinity for VEGF and produced antiangiogenic effects, suggesting its potential for treating angiogenesis-related diseases such as cancer.     

Low-dose all-trans retinoic acid enhances cytotoxicity of cisplatin and 5-fluorouracil on CD44+ cancer stem cells

Cis-diamminedichloridoplatinum(II)(CDDP)-based combination chemotherapy is frequently used in gastrointestinal cancer. The synergistic mechanism of all-trans retinoic acid (ATRA), cisplatin (CDDP) and 5-fluorouracil (5-FU) in combination remains unclear. Despite their potent antitumor properties, resistance to CDDP and 5-FU develops frequently in tumors. To clarify this mechanism, we determined the sensitivity to each drug and their combination in two gastrointestinal cancer stem cells (CSCs) subpopulation.Here, we report the identification and separation of CD44⁺ cells from human gastric carcinoma (AGS) and human esophageal squamous cell carcinoma (KYSE-30) cancer cell lines by magnetic activated cell sorting (MACS). We allowed the CD44^± cells to grow 6 days at a subtoxic concentration of ATRA and then treated with different concentration of CDDP and 5-FU for 24 h. The cytotoxicity was examined by cell proliferation MTT assay. Additionally, AO/EB staining was used for detection of apoptotic cells. In order to determine whether the growth inhibition was also associated with changes in cell cycle distribution, cell cycle analysis was performed using flow cytometry.Low concentration of ATRA (1 μM, 6days) followed by 5-FU and CDDP was found to be more effective than either drugs alone, thus resulting in synergistic cytotoxicity in Kyse-30 and AGSCD44^± cells. Furthermore, there was an indication that the combination of ATRA with 5FU and CDDP caused an increase in cell cycle arrest in G2/M and G0/G1.We conclude that low concentration of ATRA enhances the cytotoxicity of CDDP and 5FU by facilitating apoptosis and cell cycle arrest in gastrointestinal CSCs and provide a rational basis for the design of novel, well-tolerated CDDP- and 5FU-based chemotherapy in human gastrointestinal carcinoma.     

TMPRSS4 promotes invasiveness of human gastric cancer cells through activation of NF-κB/MMP-9 signaling

Transmembrane protease serine 4 (TMPRSS4) is a type-II transmembrane serine protease that is frequently upregulated in human cancers. However, little is known about the biological roles of TMPRSS4 in gastric cancer. In this study, we examined the effect of TMPRSS4 on gastric cancer cell proliferation, migration, and invasion. The expression and secretion of matrix metalloproteinase-9 (MMP-9) and activation of nuclear factor-κB (NF-κB) were determined. The involvement of NF-κB/MMP-9 signaling was checked. Our data showed that TMPRSS4 silencing significantly (P < 0.05) reduced the migration and invasion of AGS and MKN-45 gastric cancer cells, without affecting cell proliferation. Overexpression of TMPRSS4 significantly promoted cell migration and invasion. The expression and secretion of MMP-9 was significantly (P < 0.05) enhanced in TMPRSS4-overexpressing cells. TMPRSS4-overexpressing cells had a significantly (P < 0.05) lower level of IκBα and higher level of nuclear NF-κB. Luciferase reporter assay confirmed that overexpression of TMPRSS4 resulted in a 3–5-fold increase in NF-κB-dependent luciferase activity. Downregulation of MMP-9 significantly (P < 0.05) reversed the invasiveness of gastric cancer cells induced by TMPRSS4 overexpression. Moreover, pharmacological inhibition of NF-κB attenuated the invasion of TMPRSS4-overexpressing cells and the expression of MMP-9. Upregulation of TMPRSS4 enhances the invasiveness of gastric cancer cells, largely through activation of NF-κB and induction of MMP-9 expression. Our study provides the rationale for targeting TMPRSS4 in the treatment of gastric cancer.     

Cryptotanshinone induces melanoma cancer cells apoptosis via ROS-mitochondrial apoptotic pathway and impairs cell migration and invasion

Melanoma is the most serious type of skin cancer because it is highly frequency of drug resistance and can spread earlier and more quickly than other skin cancers. The objective of this research was to investigate the anticancer effects of cryptotanshinone on human melanoma cells in vitro, and explored its mechanisms of action. Our results have shown that cryptotanshinone could inhibit cell proliferation in human melanoma cell lines A2058, A375, and A875 in a dose- and time-dependent manner. In addition, flow cytometry assay showed that cryptotanshinone inhibited the proliferation of human melanoma cell line A375 by blocking cell cycle progression in G2/M phase and inducing apoptosis in a concentration-dependent manner. Moreover, western blot analysis indicated that the occurrence of its apoptosis was associated with upregulation of cleaved caspases-3 and pro-apoptotic protein Bax while downregulation of anti-apoptotic protein Bcl-2. Meanwhile, cryptotanshinone could decrease the levels of reactive oxygen species (ROS). Furthermore, cryptotanshinone also blocked A375 cell migration and invasion in vitro which was associated with the downregulation with MMP-9. Taken together, these results suggested that cryptotanshinone might be a potential drug in human melanoma treatment by inhibiting proliferation, inducing apoptosis via ROS-mitochondrial apoptotic pathway and blocking cell migration and invasion.     

Effect of resveratrol on drug resistance in colon cancer chemotherapy

To investigate the effects of resveratrol on the drug resistance of 5-FU in the colon cancer chemotherapy, an MTT assay was used to detect the effects of 5-FU and resveratrol combined with 5-FU on the proliferation of the LoVo and SW480 cell lines. Flow cytometry was used to detect the effect of 5-FU combined with resveratrol on the survival rate of the LoVo and SW480 cells. A western blot was used to detect the expression levels of the proteins associated with colon cancer. After flow sorting, the percentage of the SW480 and the LoVo cell line CD133⁺ was 97.5% and 95.8%, respectively. The cells cultured in vitro showed more rapid cell proliferation and differentiation. The MTT assay showed that as compared with the survival rate of the blank group LoVo and CD133⁺ LoVo cells, the survival rate of the cells containing the 5-FU group was lower (P < 0.05). When 5-FU was used in combination with different concentrations of resveratrol, the abovementioned phenomenon was more prominent. The sorted colon cancer cells have dry stem cells, and the sorted CD133⁺ cells are more resistant to drugs; the combination of resveratrol and 5-FU has the best effect on the colon cancer cells. Preliminary studies on the mechanism of action of the drug show that a combination of 5-FU and resveratrol regulates apoptosis in CD133⁺ colon cancer stem cells by regulating the BAX gene; however, more complex mechanisms may also be involved.

To investigate the effects of resveratrol on the drug resistance of 5-FU in the colon cancer chemotherapy, an MTT assay was used to detect the effects of 5-FU and resveratrol combined with 5-FU on the proliferation of the LoVo and SW480 cell lines.     

RNA干扰质粒对胰腺癌细胞系Panc-1原癌基因AKT2表达的影响

  目的  探讨RNA干扰质粒抑制胰腺癌细胞系Panc-1原癌基因AKT2的表达对胰腺癌细胞生长和凋亡的影响, 并初步探讨其作用机制。  方法  选择胰腺癌细胞系Panc-1, 构建特异性抑制AKT2表达的RNA干扰质粒, 瞬时和稳定转染胰腺癌细胞, 采用MTT法及软琼脂克隆形成实验检测胰腺癌细胞生长能力, Heochst染色及Annexin V-FITC/PI染色法检测细胞凋亡情况, 通过Western blot方法检测凋亡蛋白caspase-3表达; 并进行裸鼠移植瘤体内转染实验。  结果  采用RNA干扰质粒沉默胰腺癌细胞系Panc-1原癌基因AKT2, 能够有效抑制胰腺癌细胞Panc-1体外生长能力、促进细胞凋亡, 诱导凋亡激酶caspase-3的表达; 动物体内实验结果显示, 干扰质粒能够有效抑制胰腺癌细胞系Panc-1在动物体内的成瘤能力。  结论  RNA干扰质粒抑制原癌基因AKT2表达, 可有效抑制胰腺癌细胞生长, 促进凋亡, 针对原癌基因AKT2的基因治疗对胰腺癌具有重要的潜在应用价值。