Cell Apoptosis Pathways Research Articles

The Effect of Lycopene on Cancer Cell Apoptosis by Caspase-9 Concentration Measurement in Indonesian Human Prostate Cancer Cell Culture

BACKGROUND: Lycopene is an antioxidant that mostly found in daily ingredients such as tomatoes. Several studies have shown the lycopene potential in preventing prostate cancer. Nevertheless, the clinical use of lycopene as adjunctive therapy for prostate cancer is still under debate.
 AIM: The objective of the study was to determine the effect of lycopene on human prostate cancer cell culture growth by measuring caspase-9 concentration as a marker of the intrinsic pathway of apoptosis in cells.
 METHODS: This study was conducted on Indonesian prostate cancer cell culture from a patient with Gleason score 6, divided into 5 subgroups: 2 control groups and 3 treatment groups that were given 1 μM, 2 μM, and 4 μM of lycopene. Measurement of caspase-9 level was performed using enhanced chemiluminescence at 24, 28, and 72 h after lycopene addition in treatment groups. A comparative analysis was performed by two-way ANOVA.
 RESULTS: The result showed that there was a significant difference of mean caspase-9 levels in the provision of various concentrations of lycopene and time of observation (p < 0.05). Increased of mean caspase-9 levels started at 2 μM dose of lycopene at 48 h and 4 μM at 24 h (p < 0.05) and continue to rise at 72 h, but caspase-9 was not detected at 1 μM dose in every observation.
 CONCLUSION: There was a significant difference of mean caspase-9 levels in the provision of various concentrations of lycopene and time of observation.

A novel HER2-targeting antibody 5G9 identified by large-scale trastuzumab-based screening exhibits potent synergistic antitumor activity.

BackgroundPertuzumab is currently used in combination with trastuzumab as the first-line treatment for HER2-positive metastatic breast cancer. However, pertuzumab was originally developed independently from trastuzumab and was later incidentally found to have synergistic efficacy when combined with trastuzumab, it remains to be seen whether a more potent synergistic efficacy partner exists for trastuzumab.MethodsA trastuzumab-based functional assay was used to screen anti-HER2 antibodies harboring trastuzumab-synergistic antitumor activity. The lead candidate 5G9, in combination with trastuzumab, was further characterized for its bioactivities in cell proliferation, cell apoptosis, antigen-antibody endocytosis and HER2-mediated cell signaling pathway blocking. Finally, animal models were used to evaluate the in vivo synergistic antitumor efficacy of 5G9 in combination with trastuzumab.FindingsCompared to pertuzumab, 5G9 demonstrated more potent synergistic cell growth inhibitory activity when combined with trastuzumab (85% vs. 55%, P<0.001). In addition, 5G9 exhibited a higher internalization rate than pertuzumab (20% vs. 9%, P<0.05), and was able to further synergize with trastuzumab to promote antigen-antibody endocytosis. The internalization rate of the combination of 5G9 and trastuzumab was higher than that of pertuzumab and trastuzumab (35% vs. 14%, P<0.001). In vivo animal studies demonstrated that 5G9 in combination with trastuzumab showed more potent synergistic antitumor efficacy than the combination of pertuzumab and trastuzumab.Interpretation5G9, together with trastuzumab, may provide a potential opportunity for more efficacious treatment of HER2-positive cancers.FundingNational Natural Science Foundation of China; State Key Laboratory of Analytical Chemistry for Life Science.

Hepatic distribution and toxicity of zirconia nanoparticles in vivo and in vitro

Zirconia nanoparticles (ZrO2-NPs) have been increasingly used in industrial, biomedical and dental materials. Nevertheless, the scientific basis for the toxicological effects of ZrO2-NPs is poorly elucidated, and the understanding of the underlying mechanism is still limited. The hepatic biodistribution and toxicological effects of ZrO2-NPs after a single intravenous administration (20 mg/kg bw) in vivo and the toxicological mechanism toward hepatocytes in vitro were investigated. The liver showed continuous ZrO2-NP accumulation over a 28-d period. Moreover, ZrO2-NPs induced steatosis, functional injury, inflammatory response, and gene alternations in the liver. The main gene altered pathways induced by ZrO2-NP exposure were involved in metabolism, cellular processes, and human diseases. Among these pathways, lipid biosynthesis and metabolism alterations were predominant, and P53 signaling was also identified. Precious gene expression quantification further showed alterations in genes related to lipid biosynthesis and metabolism and cell apoptosis. Meanwhile, the results of the in vitro studies demonstrated ZrO2-NPs induce oxidative stress, lipid accumulation, cell apoptosis and P53-mediated signaling pathway activation in HepG2 cells. This study proves that ZrO2-NPs have impacts on the liver. There is potential concern over the hepatotoxicity of ZrO2-NPs in biomedical applications and occupational exposure through large-scale production.

Abnormal lipid metabolism induced apoptosis of spermatogenic cells by increasing testicular HSP60 protein expression.

Long-term consumption of high-fat and high-calorie foods not only causes obesity, but also may cause a decline in sperm quality in men. Rats with abnormal lipid metabolism (high-fat rats) were established by high-fat diet for 24 weeks. HE staining was used to observe the morphological changes of testis in rats, TUNEL and flow cytometer was used to detect the cell apoptosis in rat testis and in vitro. Immunohistochemistry and Western blotting were used to detect the expression of protein. After 24weeks of high-fat food feeding, the body weight, serum lipids and number of apoptotic spermatogenic cells in the high-fat group rat were significantly higher than those in the control group. In vivo, the expression of HSP60 protein in testis of high-fat rats was positive related to apoptosis of spermatogenic cells, cleaved caspase 3/caspase 3 protein expression and Bax/Bcl2 protein expression in testis of high-fat rats. Proportion of apoptotic spermatogenic cells was increased by up-regulation of HSP60 protein expression in vitro. Long-term consumption of high-fat diets can cause high expression of HSP60 and spermatogenic cells apoptosis in rats, while HSP60 over-expression promotes spermatogenic cell apoptosis and MAPK signal pathway in vitro.

Hinokitiol induces cell death and inhibits epidermal growth factor-induced cell migration and signaling pathways in human cervical adenocarcinoma

ObjectiveThe aim of this study was to examine the antitumor activity of hinokitiol for its clinical application in the treatment of human cervical carcinoma. Materials and methodsCervical carcinoma HeLa cells were treated by different concentrations of hinokitiol. Flow cytometry was used to analyze cell cycle. Senescence-associated β-galactosidase (SA-β-gal) assay was used to identify senescent cells. The effects of hinokitiol on EGF-induced cell migration were determined by wound healing and transwell migration assays. Western blot was used to detect proteins involved in cell cycle progression, apoptosis, autophagy, and EGF-induced signaling pathways. ResultsHinokitiol suppressed cell viability in a dose-dependent manner. Flow cytometric analysis indicated that hinokitiol treatment resulted in cell cycle arrest at G1 phase, with reduced number of cells in the G2/M phase. Western blot analysis further demonstrated that hinokitiol treatment increased the levels of p53 and p21, and concomitantly reduced the expression of cell cycle regulatory proteins, including cyclin D and cyclin E. SA-β-gal assay showed that hinokitiol treatment significantly induced β-galactosidase activity. In addition, treatment with hinokitiol increased the accumulation of the autophagy regulators, beclin 1 and microtubule-associated protein 1 light chain 3 (LC3-II), in a dose-dependent manner; however, it did not induce caspase-3 activation and poly ADP ribose polymerase (PARP) cleavage. In addition, epidermal growth factor-induced cell migration and c-Jun N-terminal kinase (JNK) and focal adhesion kinase (FAK) phosphorylation were significantly inhibited by hinokitiol. ConclusionOur findings revealed that hinokitiol might serve as a potential therapeutic agent for cervical carcinoma therapy.

Tandem Mass Tag-Based Quantitative Proteome Analysis of Porcine Deltacoronavirus (PDCoV)-Infected LLC Porcine Kidney Cells.

Porcine deltacoronavirus (PDCoV) is a newly emerging porcine pathogenic enteric coronavirus that can cause diarrhea, vomiting, dehydration, and a high mortality rate in piglets. At present, the understanding of PDCoV pathogenesis is very limited, which seriously hinders effective prevention and control. In this study, liquid chromatography tandem-mass spectrometry (LC–MS/MS) combined with tandem mass tag (TMT) labeling was performed to compare the differential expression of proteins in PDCoV-infected and mock-infected LLC-PK cells at 18 h post-infection (hpi). In addition, the parallel reaction monitoring (PRM) technique was used to verify the quantitative proteome data. A total of 4624 differentially expressed proteins (DEPs) were quantitated, of which 128 were significantly upregulated, and 147 were significantly downregulated. Bioinformatics analysis revealed that these DEPs were involved mainly in the defense response, apoptosis, and the immune system, and several DEPs may be related to interferon-stimulated genes and the immune system. Based on DEP bioinformatics analysis, we propose that PDCoV infection may utilize the apoptosis pathway of host cells to achieve maximum viral replication. Meanwhile, the host may be able to stimulate the transcription of interferon-stimulated genes (ISGs) through the JAK/STAT signaling pathway to resist the virus. Overall, in this study, we presented the first application of proteomics analysis to determine the protein profile of PDCoV-infected cells, which provides valuable information with respect to better understanding the host response to PDCoV infection and the specific pathogenesis of PDCoV infection.

A novel antitumor protein from the mushroom Pholiota nameko induces apoptosis of human breast adenocarcinoma MCF-7 cells in vivo and modulates cytokine secretion in mice bearing MCF-7 xenografts

Breast cancer is the most common malignant tumor in women and one of the three most common cancers worldwide. It is a life-threatening disease among women and the leading cause of death among women. New drugs or new drug translations and laboratory clinical studies are ongoing. A new antitumor protein (PNAP) purified from edible fungus Pholiota nameko has potential for treating breast cancer. We have previously found that PNAP exhibits anti-proliferative and apoptosis-inducing activities in a human breast cancer cell line (MCF-7). In this study, we constructed a BALB/c mouse model of MCF-7 tumor xenografts. In vivo experiments show that PNAP can effectively inhibit the malignant proliferation of MCF-7 solid tumors. This is because PNAP can successfully activate the death receptor pathway and mitochondrial apoptosis pathway of MCF-7 tumor cells in vivo, and induce tumor cells to wither. It is estimated that PNAP may also have an immunoregulatory ability to indirectly inhibit malignant proliferation of tumors. We also found that PNAP may also have the immunomodulatory ability to indirectly inhibit the malignant proliferation of tumors, which can shift the balance of Th1/Th2 to Th1 and eventually inhibit the growth of tumors. The study reveals a new therapeutic approach for breast cancer patient.

Necroptosis restricts influenza A virus as a stand-alone cell death mechanism.

Influenza A virus (IAV) activates ZBP1-initiated RIPK3-dependent parallel pathways of necroptosis and apoptosis in infected cells. Although mice deficient in both pathways fail to control IAV and succumb to lethal respiratory infection, RIPK3-mediated apoptosis by itself can limit IAV, without need for necroptosis. However, whether necroptosis, conventionally considered a fail-safe cell death mechanism to apoptosis, can restrict IAV-or indeed any virus-in the absence of apoptosis is not known. Here, we use mice selectively deficient in IAV-activated apoptosis to show that necroptosis drives robust antiviral immune responses and promotes effective virus clearance from infected lungs when apoptosis is absent. We also demonstrate that apoptosis and necroptosis are mutually exclusive fates in IAV-infected cells. Thus, necroptosis is an independent, "stand-alone" cell death mechanism that fully compensates for the absence of apoptosis in antiviral host defense.

Abstract 234: A novel, small molecule inhibitor of dihydroorotate dehydrogenase (DHODH), RP7214, potentiates activity of chemotherapeutics in breast and colorectal cancers

Abstract Background: Dihydroorotate dehydrogenase (DHODH) is an enzyme which is critically involved in process of de novo pyrimidine biosynthesis. Therefore, it plays important roles in cell proliferation, mitosis and cellular metabolism. Importantly, overexpression of DHODH has been found in various types of malignant tumors including melanoma, myeloma, and lymphoma. Moreover, from Oncomine database, we observed that colorectal and kidney cancers and lymphoma have much higher expression of DHODH compared to normal cells. These findings suggest that DHODH could be a promising target for cancer therapy. Recently, we have designed and synthesized a novel small molecule inhibitor (RP7214) of DHODH and found that it has potent inhibitory effects in acute myeloid leukemia. However, the inhibitory effect of RP7214 on solid tumor-derived cell lines and the downstream oncogenic markers is not completely understood. Methods: In vitro studies aimed at elucidating the effects of RP7214 on cell proliferation and the underlying molecular mechanisms were conducted using breast adenocarcinoma (MDA-MB-231) and colorectal carcinoma cell lines (HT-29 and HCT-116). We also investigated whether RP7214 could potentiate the anti-cancer activities of proven chemotherapeutics such as Verzenio, Doxorubicin, and Oxaliplatin, for the treatment of breast and colorectal cancers. Results: RP7214 significantly (P&amp;lt;0.05) inhibited proliferation and suppressed colony formation in MDA-MB-231, HT-29, and HCT-116 cells. Isobologram analysis showed that RP7214 synergistically enhanced the inhibitory effects of Verzenio and Doxorubicin on MDA-MB-231 cells proliferation. RP7214 and Oxaliplatin also showed synergistic inhibitory effects on the growth of HT-29 and HCT-116 colorectal cancer cells. Up-regulation of p21 with concomitant down-regulation of Bcl-2, myc, CDK6, and UAP1 in MDA-MB-231 cells treated with RP7214 alone or in combination with Doxorubicin was observed. Similarly, RP7214 also up-regulated the expression of p21 and Bax, and down-regulated the expression of CDK4, Akt, HOXA9, EZH2, and snail in HT-29 or HCT-116 cells treated with or without Oxaliplatin. These results suggest that the inhibitory effects of RP7214 on cancer cells could be mediated through the regulation of cell cycle, apoptosis, and metabolism pathways. Conclusions: Based on these results, we conclude that RP7214 could be a promising DHODH inhibitor used in combination with conventional chemotherapeutics for better treatment of breast and colorectal cancers. We are now conducting animal experiment to confirm the value of RP7214 in vivo in combination treatment of breast and colorectal cancers. Citation Format: Asfar S. Azmi, Amro Aboukameel, Mohammad Najeeb Al-Hallak, Philip A. Philip, Kumar Penmetsa, Srikant Visvanadha, Gabriel Mpilla, Rachel Sexton, Ramzi M. Mohammad. A novel, small molecule inhibitor of dihydroorotate dehydrogenase (DHODH), RP7214, potentiates activity of chemotherapeutics in breast and colorectal cancers [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 234.

Abstract 2512: Pan-cancer characterization of the tumor and immune microenvironment facilitates identification of cancer-specific biological signatures

Abstract Background: A better understanding of the characteristics of cancer across different indications is required to drive the development of personalized treatments, inform therapy decisions, and improve outcomes. Integrating data from the tumor and the immune system can enable the identification of comprehensive biological signatures and composite biomarkers for the improved stratification of responders/progressors. Here, we describe a pan cancer study, including an enhanced whole-exome and transcriptome sequencing approach, across over 500 samples representing 13 tumor types, analyzed at high depth using the ImmunoID NeXT platform. Methods: We sequenced paired tumor-normal samples on the ImmunoID NeXT platform, an enhanced exome/transcriptome-based diagnostic platform that can simultaneously profile the tumor and immune microenvironment from a single FFPE sample, across all of the approximately 20,000 genes. For each sample, we analyzed a broad set of features focused on both the tumor and immune system. From DNA, we profiled small variants, CNAs, MSI status, oncoviruses, HLA LOH, and neoantigens. From RNA, we profiled gene expression, small variants, fusions, TILs, TCR, BCR, and immune signatures. Integrated analyses assessing the impact of each feature, both within and across tumor types, were performed across the cohort. Results: Through immunogenomic analysis we identified striking differences in both tumor and TME profiles across cancer types. In addition to mutation and neoantigen burden, by. we also computed a composite neoantigen score for each sample, which we have shown in a separate melanoma study can be a stronger predictor of response to immunotherapy. The composite neoantigen score integrates neoantigen prediction with mechanisms of tumor escape that can affect neoantigen presentation, providing a more accurate model of the antigen presentation biology. We also looked at the distribution of HLA LOH using our DASH algorithm and found differences in LOH frequency between tumor types. For example, we found HLA LOH to be five timesmore common in lung cancer than breast cancer. Further, we profiled immune gene signatures, including Gejewski and Ribas signatures, highlighting varied immune activation across cancer types. Analysis of somatic alterations in pathways controlling cell growth, PI3K/AKT signaling, apoptosis, and other canonical pathways revealed malignancy-specific alteration frequencies. The varying frequency, and combination of these alterations is indicative of a complex hierarchy of cross-talk between pathways, which operates in a cancer specific manner. Conclusions: We performed a broad integrated analysis of the tumor and immune microenvironment for over 500 samples across 13 different tumor types using the ImmunoID NeXT platform. This comprehensive profiling revealed significant differences between cancer types beyond mutational burden, including neoantigen burden, immune microenvironment differences, and incidence of putative tumor escape mechanisms Citation Format: Sean Michael Boyle, Charles Abbott, Eric Levy, Rachel Marty Pyke, Dattatreya Mellacheruvu, Simo Zhang, Mengyao Tan, Rena McClory, John West, Richard Chen. Pan-cancer characterization of the tumor and immune microenvironment facilitates identification of cancer-specific biological signatures [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2512.

In Vitro Anticancer Effects of Stilbene Derivatives: Mechanistic Studies on HeLa and MCF-7 Cells.

The growing prevalence of cancer and the resulting chemoresistance exert a huge burden on healthcare systems and impose a great challenge to public health around the world. In efforts to develop new chemotherapeutic agents for cancer treatment, a class of heterocyclic compounds i.e. triazine-based molecules were investigated as anticancer agents. New triazine hybrids of stilbene were synthesized and evaluated as anticancer agents for cervical (HeLa) and breast (MCF-7) carcinoma cells. The compound (7e), sodium (E)-6,6'-(ethene-1,2- diyl)bis(3-((4-chloro-6-((3-luorophenyl)amino)-1,3,5-triazin-2-yl)amino)benzenesulfonate) was found to be most potent among synthesized derivatives and was explored further for detailed mechanistic studies. In a set comprised of twelve derivatives, compound 7e, sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((4- chloro-6-((3-luorophenyl)amino)-1,3,5-triazin-2-yl)amino)benzenesulfonate) was found most potent inhibitor for HeLa and MCF-7 cells. The present study has revealed that compound 7e may activate mitochondrial pathway of apoptosis in HeLa and MCF-7 cells which was assessed by DNA binding studies, estimation of the release of Lactate Dehydrogenase (LDH), fluorescence imaging, production of Reactive Oxygen Species (ROS) in cancer cells, analysis of cell cycle by flow cytometry, change in Mitochondrial Membrane Potential (MMP) and activation of caspase-9 and caspase-3. Compound 7e may serve as a lead in designing new anticancer compounds based on stilbene scaffold.

Mitochondrion-Directed Nanoparticles Loaded with a Natural Compound and a microRNA for Promoting Cancer Cell Death via the Modulation of Tumor Metabolism and Mitochondrial Dynamics.

Mitochondrial dysfunction may cause cancer and metabolic syndrome. Ellagic acid (abbreviated as E), a phytochemical, possesses anticancer activity. MicroRNA 125 (miR-125) may regulate metabolism. However, E has low aqueous solubility, and miR-125 is unstable in a biological fluid. Hence, this study aimed to develop nanoparticle formulations for the co-treatment of miR-125 and E. These nanoparticles were modified with one mitochondrion-directed peptide and a tumor-targeted ligand, and their modulating effects on mitochondrial dysfunction, antitumor efficacy, and safety in head and neck cancer (HNC) were evaluated. Results revealed that miR-125- and E-loaded nanoparticles effectively targeted cancer cells and intracellular mitochondria. The co-treatment significantly altered cellular bioenergetics, lipid, and glucose metabolism in human tongue squamous carcinoma SAS cells. This combination therapy also regulated protein expression associated with bioenergenesis and mitochondrial dynamics. These formulations also modulated multiple pathways of tumor metabolism, apoptosis, resistance, and metastasis in SAS cells. In vivo mouse experiments showed that the combined treatment of miR-125 and E nanoparticles exhibited significant hypoglycemic and hypolipidemic effects. The combinatorial therapy of E and miR-125 nanoparticles effectively reduced SAS tumor growth. To our best knowledge, this prospective study provided a basis for combining miRNA with a natural compound in nanoformulations to regulate mitochondrial dysfunction and energy metabolism associated with cancer.

Transcriptomic analysis reveals differential gene expression, alternative splicing, and novel exons during mouse trophoblast stem cell differentiation

BackgroundDifferentiation of mouse trophoblast stem cells (TSCs) to trophoblast giant cells (TGCs) has been widely used as a model system to study placental development and function. While several differentially expressed genes, including regulators of TSC differentiation, have been identified, a comprehensive analysis of the global expression of genes and splice variants in the two cell types has not been reported.ResultsHere, we report ~ 7800 differentially expressed genes in TGCs compared to TSCs which include regulators of the cell cycle, apoptosis, cytoskeleton, cell mobility, embryo implantation, metabolism, and various signaling pathways. We show that several mitotic proteins, including Aurora A kinase, were downregulated in TGCs and that the activity of Aurora A kinase is required for the maintenance of TSCs. We also identify hitherto undiscovered, cell-type specific alternative splicing events in 31 genes in the two cell types. Finally, we also report 19 novel exons in 12 genes which are expressed in both TSCs and TGCs.ConclusionsOverall, our results uncover several potential regulators of TSC differentiation and TGC function, thereby providing a valuable resource for developmental and molecular biologists interested in the study of stem cell differentiation and embryonic development.

Protective effect of Lycium barbarum polysaccharide on ethanol-induced injury in human hepatocyte and its mechanism.

The purpose of this research is to study the effect of Lycium barbarum polysaccharide on ethanol-induced liver injury and its mechanism. The cell survival rate, the apoptosis rate, and the intracellular ROS level was detected by MTT assay, flow cytometry, laser confocal microscopy, and fluorescence spectrophotometry, respectively. The antioxidativeindices were determined by ELISA kits and the protein level was detected by western blot. The result showed Lycium barbarum polysaccharide could protect ethanol-induced cell injury by reducing cell apoptosis and regulating the levels of indicators related to oxidative stress, such as ROS, MDA, SOD, etc. In addition, LBP could increase the nuclear expression of Nrf2 protein and significantly up-regulate the expression levels of Nrf2 protein and its downstream proteins, such as HO-1, NQO1, and GCLC in the cell nucleus. Therefore, Lycium barbarum polysaccharide has a protective effect on ethanol-induced liver cell injury and it plays the role in cell apoptosis pathway and oxidative stress pathway. PRACTICAL APPLICATIONS: Lycium barbarum is a kind of food that can be used as food and medicine in China. The result showed that Lycium barbarum polysaccharide could protect ethanol-induced liver cell injury, which is beneficial to the application of LBP in functional food.

Antipsychotics drug aripiprazole as a lead against breast cancer cell line (MCF-7) in vitro.

Breast cancer is the second leading cause of death among women globally. The existing treatment options for breast cancer are largely associated with severe toxicities, and lower efficacies. Therefore, there is an urgent need for the development of non-toxic effective drugs against breast cancer. For this purpose, drug repositioning strategy was used to evaluate the anti-cancer potential of a library of heterocyclic drugs. The major advantage of drug repurposing is that the pharmacokinetic, pharmacodynamic, and toxicity profiles of drugs are well documented. In the current study, we screened 97 drugs of different chemical classes, and among them aripiprazole, an antipsychotic drug, was found to be sufficiently active against breast cancer cell line MCF-7. Aripiprazole showed a cytotoxicity (IC50 = 12.1 ± 0.40 μM) to MCF-7 cells, comparable to the standard anticancer drug doxorubicin (IC50 = 1.25 ± 0.34 μM). Aripiprazole was also found to be active against other cancer cell lines, including MDA-MB-231 (IC50 = 19.83 ± 0.27 μM), AU565 (IC50 = 18.02 ± 0.44 μM), and BT-474 (IC50 = 36.42 ± 0.12 μM). Aripiprazole significantly inhibited the cell cycle progression at subG0G1 phase, and enhanced apoptosis in MCF-7 breast cancer cells. The drug was also able to significantly increase the nuclear condensation, and modulated the expression of certain genes involved in breast cancer, such as caspases 3, and 9, BAK-1, C-MYC, BCL2L1, BCL-10, and BCL-2. Further studies are needed to explore the effect of aripiprazole on intrinsic and extrinsic pathways of apoptosis in cancer cells.

Knockdown of miR-183 Enhances the Cisplatin-Induced Apoptosis in Esophageal Cancer Through Increase of FOXO1 Expression.

BackgroundAs an important member of platinum-based chemotherapeutic drugs, cisplatin is effective and is commonly used in the treatment of esophageal cancer. However, repeated use of cisplatin usually causes severe side-effects on patients. Novel approaches should be explored to increase the sensitivity of cancer cells to cisplatin.MethodsThe expression level of miR-183 in esophageal cancer tissues and cell lines was measured by quantitative reverse transcriptase real-time PCR (qRT-PCR). The sensitivity of EC cell lines to cisplatin was evaluated by CCK-8 assay and flow cytometry. Luciferase reporter assay was used to confirm the association between miR-183 and FOXO1. The apoptosis pathway of EC cells was tested by Western blot assay.ResultsThe expression level of miR-183 was increased in esophageal cancer patients’ tumor tissues and esophageal cancer cell lines. However, knockdown of miR-183 was found to enhance the effect of cisplatin on inducing the apoptotic cell death of esophageal cancer cells. In the mechanism research, we proved that FOXO1 was the target of miR-183 in esophageal cancer cells. Inhibition of miR-183 increased the expression of FOXO1 to promote the expression of Bim and Noxa. As Bim and Noxa acted as key pro-apoptotic proteins in mitochondrial apoptosis, inhibition of miR-183 enhanced the cisplatin-induced apoptosis pathway in esophageal cancer.ConclusionKnockdown of miR-183 enhanced the cisplatin-induced apoptosis in esophageal cancer through an increase of FOXO1 expression.

Correlation of Blimp1 with ATF4/CHOP Signaling Pathway in Multiple Myeloma U266 Cells

To investigate whether Blimp1 plays an anti-apoptosis role in myeloma by interfering with ATF4/CHOP cell apoptosis pathway induced by endoplasmic reticulum stress, and to explore the anti-myeloma mechanism of aspirin. The bone marrow fluid of 40 newly diagnosed multiple myeloma patients without treatment and 30 control people with relatively normal bone marrow was collected. Flow cytometry was used to separated the normal and abnormal plasma cells, LV-Blimp1-RNAi (40051-2) recombinant lentivirus down-regulates the expression of Blimp-1 in U266 cell line and detected the changes of the expression of ATF4 and CHOP gene. U266 cells were stimulated by aspirin at different concentrations (0, 0.5, 2.5, 5.0 mmol/L) in vitro. Then the effect of aspirin on proliferation of U266 cells was measured by CCK-8 assay, the mRNA expression levels of Blimp1, ATF4 and CHOP in four groups were detected by real-time PCR. The expression level of Blimp1 in phenotype abnormal plasma cells was significantly increased as compared with normal cells, while the expression of ATF4 and CHOP in phenotype abnormal plasma cells was significantly decreased as compared with normal cells (P＜0.05). In the case of MOI=100, the transfection efficiency of U266 cells was beyond 80% as detected by fluorescence microscopy. Compared with blank conrol and negatine control groups, Blimp1 mRNA expression level in positive group was significantly reduced while ATF4 and CHOP expression significantly increased. CCK-8 showed that the proliferation activity of U266 cells could be inhibited by aspirin, which showed a time-and dose-dependent manner; at the same time, the expression level of Blimp1 in U266 cells were decreased with the increasing of aspirin concentration, while the expression level of ATF4 and CHOP was increased with the increasing of aspirin concentration. Blimp1 may display the anti-apoptosis of myeloma cells through interfering with ATF4/CHOP signaling pathway; low dose of aspirin may play anti-myeloma effect by inhibiting the expression of Blimp1 in myeloma cells.

Viral interactions with non-homologous end-joining: a game of hide-and-seek.

There are extensive interactions between viruses and the host DNA damage response (DDR) machinery. The outcome of these interactions includes not only direct effects on viral nucleic acids and genome replication, but also the activation of host stress response signalling pathways that can have further, indirect effects on viral life cycles. The non-homologous end-joining (NHEJ) pathway is responsible for the rapid and imprecise repair of DNA double-stranded breaks in the nucleus that would otherwise be highly toxic. Whilst directly repairing DNA, components of the NHEJ machinery, in particular the DNA-dependent protein kinase (DNA-PK), can activate a raft of downstream signalling events that activate antiviral, cell cycle checkpoint and apoptosis pathways. This combination of possible outcomes results in NHEJ being pro- or antiviral depending on the infection. In this review we will describe the broad range of interactions between NHEJ components and viruses and their consequences for both host and pathogen.

The effect of compound kushen injection on cancer cells: Integrated identification of candidate molecular mechanisms.

Traditional Chinese Medicine (TCM) preparations are often extracts of single or multiple herbs containing hundreds of compounds, and hence it has been difficult to study their mechanisms of action. Compound Kushen Injection (CKI) is a complex mixture of compounds extracted from two medicinal plants and has been used in Chinese hospitals to treat cancer for over twenty years. To demonstrate that a systematic analysis of molecular changes resulting from complex mixtures of bioactives from TCM can identify a core set of differentially expressed (DE) genes and a reproducible set of candidate pathways. We used in vitro cancer models to measure the effect of CKI on cell cycle phases and apoptosis, and correlated those phenotypes with CKI induced changes in gene expression. We treated two cancer cell lines with or without CKI and assessed the resulting phenotypes by employing cell viability and proliferation assays. Based on these results, we carried out high-throughput transcriptome data analysis to identify genes and candidate pathways perturbed by CKI. We integrated these differential gene expression results with previously reported results and carried out validation of selected differentially expressed genes. CKI induced cell-cycle arrest and apoptosis in the cancer cell lines tested. In these cells CKI also altered the expression of 363 core candidate genes associated with cell cycle, apoptosis, DNA replication/repair, and various cancer pathways. Of these, 7 are clinically relevant to cancer diagnosis or therapy, 14 are cell cycle regulators, and most of these 21 candidates are downregulated by CKI. Comparison of our core candidate genes to a database of plant medicinal compounds and their effects on gene expression identified one-to-one, one-to-many and many-to-many regulatory relationships between compounds in CKI and DE genes. By identifying genes and promising candidate pathways associated with CKI treatment based on our transcriptome-based analysis, we have shown that this approach is useful for the systematic analysis of molecular changes resulting from complex mixtures of bioactives.

Nuclear Factor-κB Pathway Mediates the Molecular Pathogenesis of LMNA-Related Muscular Dystrophies.

LMNA-related muscular dystrophies are caused by mutations of the LMNA gene. Inflammatory changes and cellular apoptosis are significant pathological findings in the muscle cells of these patients. We aimed to investigate the roles of nuclear factor-κB (NF-κB) mediated inflammation as a molecular mechanism for the pathogenesis of LMNA-related muscular dystrophies. Muscle specimen of a patient with LMNA gene mutation (c.1117A>G, p.I373V, reported in our previous work) showed significant inflammatory changes. The ultrastructure of muscle cells showed severe nuclear abnormalities compared with the control. Therefore, we used this mutation to establish mutant cell line for in vitro studies. Transfected human embryonic kidney 293 (HEK293) cells containing a mutant construct from this patient showed irregular nuclear morphology. Mass spectrometry analysis suggested genomic instability and augmented expression of apoptosis-related genes. We detected activation of NF-κB pathway in LMNA mutant cells which promoted the expression of downstream inflammatory factors. The LMNA mutation also activated the molecular pathway of apoptosis in LMNA mutant cells. These are important molecular mechanisms underlying the pathogenesis of LMNA-related muscular dystrophies. Our research provides crucial evidence for future pathogenetic studies and possible treatment strategies for LMNA-related muscular dystrophies.