Apoptosis-inducing Effects Research Articles

Exploring the Anticancer Efficacy of Mixture of Local Probiotics on MDA-MB-231 and MCF-7 Breast Cell Lines

This study aimed to investigate the cytotoxicity of a probiotic mixture on human breast cancer cell lines. To prepare the mixture, local probiotic bacteria were cultured, and the lyophilized supernatant was applied for downstream experiments. The antioxidant activity, total phenol content (TPC), and fatty acid composition of the bacterial supernatant (BS) were also measured. The possible cytotoxic/anti-proliferative effect of the probiotic mixture was accessed on both breast cancer cell lines at different concentrations using MTT assay. Furthermore, the apoptosis-inducing effects of the same mixture was studied by DAPI staining. The highest level of antioxidant activity and total phenol content (TPC) were detected for the BS at 3200 µg/ml. According to the GC–MS analysis, linoleic acid (37.40 %) and oleic acid (26.93 %) were identified as the major fatty acids of the BS. The MTT assay showed that the BS has anti-proliferative effects on MDA-MB-231 and MCF-7 cells in a time- and dose-dependent manner (IC50: 3200 μg/ml). The apoptosis-inducing effects of the mixture was confirmed in both cell lines through morphological analyses of the cells’ nucleoli, and the formation of apoptotic bodies. According to these experiments, cytotoxic effects and apoptosis-inducing potential were confirmed for the BS against two human breast cancer cell lines, including MDA-MB-231, and MCF-7. Hence, it could be considered as a suitable anti-cancer agent.

Digoxin reduces the incidence of prostate cancer but increases the cancer-specific mortality: A systematic review and pooled analysis.

Digoxin, a commonly used drug for congestive heart failure and cardiac arrhythmias, has been reported to exert cytotoxic and apoptosis-inducing effects on prostate cancer (PCa) cells. In this study, we aimed to perform a pooled analysis to summarise all the evidence related to the effects of digoxin on PCa development. Four electronic databases were systematically searched to filter the eligible studies. The hazard ratio (HR) with its 95% confidence interval (CI) was calculated. This study was registered on PROSPERO (ID: CRD42021226885). Ten clinical studies with a total of 108,444 participants (15,835 individuals were digoxin users) were included. The pooled result from 6 included studies demonstrated that digoxin usage was correlated with a significant decrease in PCa risk (adjusted RR=0.892, 95% CI: 0.799-0.997, p=.044) when compared with the nonusers. Synthetic result of 4 eligible studies revealed that digoxin significantly correlated with higher prostate cancer-specific mortality than the controls (adjusted HR=1.142, 95% CI: 1.005-1.297). No statistical heterogeneity was detected during this analysis (all I2 <50%, p>.1). Our study confirmed a preventive effect of digoxin usage for the risk of PCa in men. However, digoxin use was associated with a significantly elevated risk of prostate cancer-specific mortality. This finding needs more well-designed studies to better interpret the causality.

Efficacy of green synthesized silver nanoparticles via ginger rhizome extract against Leishmania major in vitro.

IntroductionLeishmaniasis is a major public health problem that causes by parasite of the genus Leishmania. The pentavalent antimonial compounds that used for treatment are not safe or effective enough. The aim of the present study was preparation and evaluation of the efficacy of green synthesized silver nanoparticles against Leishmania major (L. major) in vitro.MethodsTo synthesis silver (Ag) nanoparticles (NPs), ginger extract was added to the 0.2mM AgNO3 aqueous solution (1:20). Effects of different concentrations of Ag-NPs on the number of L. major promastigotes were investigated using counting assay. The MTT test was applied to determine the toxicity of Ag-NPs on promastigotes of L. major, as well as, macrophage cells. Then, to evaluate the anti-amastigotes effects of Ag-NPs, parasites within the macrophages were counted by light microscope. Furthermore, to determine the induced apoptosis and necrotic effects of Ag-NPs on promastigotes, flow cytometry method was employed using annexin staining.ResultsThe effect of Ag-NPs on promastigotes and amastigotes of L. major was effective and has a reverse relationship with its concentration. According to the results of anti-amastigote assay, the IC50 value of this nanoparticle was estimated 2.35 ppm after 72h. Also, Ag-NPs caused Programmed Cell Death (PCD) in promastigotes of L. major and showed 60.18% of apoptosis.DiscussionBased on the mentioned results, it can be concluded that Ag NPs has a beneficial effect on promastigote and amastigote forms of L. major in vitro. Hence, these nanoparticles could be applied as promising antileishmanial agents for treatment of Leishmania infections.

Silver Nanoparticles Derived by Artemisiaarborescens Reveal Anticancer and Apoptosis-Inducing Effects.

The fight against cancer is one of the main challenges for medical research. Recently, nanotechnology has made significant progress, providing possibilities for developing innovative nanomaterials to overcome the common limitations of current therapies. In this context, silver nanoparticles (AgNPs) represent a promising nano-tool able to offer interesting applications for cancer research. Following this path, we combined the silver proprieties with Artemisia arborescens characteristics, producing novel nanoparticles called Artemisia–AgNPs. A “green” synthesis method was performed to produce Artemisia–AgNPs, using Artemisia arborescens extracts. This kind of photosynthesis is an eco-friendly, inexpensive, and fast approach. Moreover, the bioorganic molecules of plant extracts improved the biocompatibility and efficacy of Artemisia–AgNPs. The Artemisia–AgNPs were fully characterized and tested to compare their effects on various cancer cell lines, in particular HeLa and MCF-7. Artemisia–AgNPs treatment showed dose-dependent growth inhibition of cancer cells. Moreover, we evaluated their impact on the cell cycle, observing a G1 arrest mediated by Artemisia–AgNPs treatment. Using a clonogenic assay after treatment, we observed a complete lack of cell colonies, which demonstrated cell reproducibility death. To have a broader overview on gene expression impact, we performed RNA-sequencing, which demonstrated the potential of Artemisia–AgNPs as a suitable candidate tool in cancer research.

Titania Nanosheet Generates Peroxynitrite-Dependent S-Nitrosylation and Enhances p53 Function in Lung Cancer Cells.

Metal nanomaterials can enhance the efficacy of current cancer therapies. Here, we show that Ti0.8O2 nanosheets cause cytotoxicity in several lung cancer cells but not in normal cells. The nanosheet-treated cells showed certain apoptosis characteristics. Protein analysis further indicated the activation of the p53-dependent death mechanism. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analyses revealed the cellular uptake of the nanosheets and the induction of cell morphological change. The nanosheets also exhibited a substantial apoptosis effect on drug-resistant metastatic primary lung cancer cells, and it was found that the potency of the nanosheets was dramatically higher than standard drugs. Ti0.8O2 nanosheets induce apoptosis through a molecular mechanism involving peroxynitrite (ONOO−) generation. As peroxynitrite is known to be a potent inducer of S-nitrosylation, we further found that the nanosheets mediated the S-nitrosylation of p53 at C182, resulting in higher protein-protein complex stability, and this was likely to induce the surrounding residues, located in the interface region, to bind more strongly to each other. Molecular dynamics analysis revealed that S-nitrosylation stabilized the p53 dimer with a of <−1.5 kcal/mol. These results provide novel insight on the apoptosis induction effect of the nanosheets via a molecular mechanism involving S-nitrosylation of the p53 protein, emphasizing the mechanism of action of nanomaterials for cancer therapy.

Cytotoxicity and Apoptosis Inducing Effects of Some Lathyrane and Tigliane Diterpenes against Breast Cancer Cell Lines

Natural compounds and especially herbal medicine are of great interest due to their various biological effects and their potential to act as a drug for the treatment of various neoplasms especially breast cancer that we are facing with its increasing prevalence around the world. The aim of this study was to evaluate the cytotoxic and cell death mechanism of some diterpenoids (Lathyrane or Tigliane) extracted from the Euphorbia sogdiana Popov against two breast cancer cell lines, MCF-7 and 4T1. Determination the cytotoxic effects of four various diterpenoids was performed using MTT assay against MCF-7, 4T1, and HUVEC cell lines. The IC50 of each compound against cell lines was determined by drawing the dose-response graph using graph Pad prism software. Finally, the apoptotic effects of compound with the most cytotoxic effects was determined by flow cytometry assay for 24 hrs of incubation in IC50 concentration. Statistical analysis confirmed compound (3) with the most cytotoxicity against both cancer cell lines. The IC50 of compound (3) was determined as 10.1 ± 5, 28 ± 5, and 50 ± 3 µg/ml, for MCF-7, 4T1, and HUVEC cells, respectively. Furthermore, the cells treated with 5 and 10 μg/ml of compound (3) for 24 hrs, showed 49 and 57% of apoptosis. These surveyed compounds have the potential to be considered as useful anti-breast cancer agents due to the great cytotoxicity and apoptotic effects against related cancer cell lines and safety profile according to their rational selectivity index.

The Association of Cell Surface Fibromodulin Expression and Bladder Carcinoma.

Fibromodulin (FMOD) is a secretory protein which is considered a major component of extracellular matrix. Its dysregulation in different types of cancer implies it as a promising target for cancer therapy. Within the scope of its rather wide expression in different tumors, we studied expression of FMOD and effect of anti-FMOD antibody in bladder cancer cells in order to identify new target for diagnostic and therapeutic interventions. We report here for the first time the expression of FMOD in bladder cancer cell lines in comparison to the normal cell line and tissues. A peptide-based produced anti-FMOD murine monoclonal antibody (mAb) (clone 2C2-A1) was applied for evaluation of FMOD expression in bladder cancer and normal tissues by immunohistochemistry (IHC) staining. Furthermore, the expression of FMOD was examined in human bladder cell lines, 5637 and EJ138, as well as a non-cancerous human cell line, human fetal foreskin fibroblast (HFFF), by immunocytochemistry (ICC) and flow cytometry. The apoptosis induction of anti-FMOD mAb was also evaluated in bladder cancer cells. IHC and ICC analyses revealed that the qualitative expression of FMOD in bladder cancer tissues and cell lines is higher than in normal tissues and cell lines. Flow cytometry analyses revealed that 2C2-A1 mAb could recognize FMOD expression in 84.05 ± 1.85%, 46.1 ± .4% , and 2.56 ± 1.26% of 5637, EJ138, and HFFF cells, respectively. An effective apoptosis induction was detected in 5637 and EJ138 cells with no significant effect on HFFF cell. To our knowledge, this is for the first time reporting surface expression of FMOD in bladder cancer. This significant surface expression of FMOD in bladder cancer with no expression in normal bladder tissues and the capacity of inducing apoptosis through directed targeting of FMOD with specific monoclonal antibody might candidates FMOD as a diagnostic marker as well as a potential immunotargeting with monoclonal antibody.

Abstract 1239: Putative Bax activators GL0385 and GL0388 for targeted breast cancer therapy

Abstract Breast cancer is one of the leading causes of cancer-related death in American women and the most common cancer in women with more than 270,000 estimated new cases in 2020 in the US. This is about 30% of the estimated total cancer patients in American women and therefore a major threat to the society with an urgent need of advanced treatment regimens. Especially the subgroup of estrogen receptor (ER)-negative breast cancers (ENBC), which are approximately one quarter to a third of all breast cancer, are lacking efficient treatment methods. Particularly dangerous is triple-negative breast cancer (TNBC), a subtype of ENBC with a high metastatic potential, since 90% of cancer-related deaths are due to metastasis. However, an effective drug for ENBC/TNCB treatment remains to be developed. The pro-apoptotic protein Bax was shown to be downregulated in most cancers. As a result, the balance between Bax and its antagonist Bcl-2 gets disturbed which leads to dysfunctional apoptotic signaling in favor of cancer cell survival. Upon activation, Bax is translocated from the cytosol to the mitochondria where it binds to the outer mitochondrial membrane. This leads to increased permeability of the mitochondrial membrane and a release of pro-apoptosis related molecules such as cytochrome C and others. Hence, drug discovery for Bax activation is a promising approach for developing novel cancer therapies. The small molecules GL0385 and GL0388 were identified from our progenitor molecules including small-molecule Bax activator (SMBA1), CYD-2-11, and CYD-4-61. The CYD compounds specifically target the Ser184 residue of Bax and have shown apoptosis-inducing effects both in vitro and in vivo. These lead Bax activators provided proof-of-concept studies while further optimization is needed to mitigate potential toxicity, leading to the discovery of advanced lead molecules GL0385 and GL0388. Treatment with either GL0385 or GL0388 resulted in significantly decreased cell proliferation in MDA-MB-231, MDA-MB-468, BT549, MDA-MB-453, MCF-7, and MDA-MB-361 cells. Additionally, MDA-MB-231 cells treated with GL0385 or GL0388 showed reduced migration and invasion, compared to control cells. Western blot analysis confirmed increased protein levels of Bax, cleaved PARP, cleaved caspase 3, and cytochrome C. These results show that GL0385 and GL388 have a significant impact on cancer cell proliferation and cellular motility which are main attributes of metastasizing cells. Additionally, increased expression of apoptosis markers indicates that these compounds may be beneficial to regain the anti-apoptotic/pro-apoptotic balance in breast cancer cells. Therefore, GL0385 and GL0388 are promising new advanced lead compounds towards the development of effective therapies for breast cancers and other cancers. Further investigation is warranted to fully explore their anti-cancer potential and mechanisms of action. Citation Format: Doerte R. Fricke, Hyejin Kim, Gang Liu, Haiying Chen, Jia Zhou, Qiang Shen. Putative Bax activators GL0385 and GL0388 for targeted breast cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1239.

Forsythia suspensa extract has inhibitory effect on proliferation and apoptosis of A549 lung cancer cells

Purpose: To investigate the effect of Forsythia suspensa extract (FSE) on apoptosis and proliferation in A549 human lung cancer cells.&#x0D; Methods: Inverted microscope was employed to observe morphological changes in A549 cells after exposure to FSE. Trypan blue staining of living cells was used to construct the cell growth curve after treatment with varying concentrations of FSE. The influence of FSE on cell proliferation, apoptosis and cell cycle was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, while protein expressions of key apoptosis-related enzymes were evaluated by immunocytochemical method.&#x0D; Results: FSE inhibited the growth of A549 lung cancer cells at a concentration range of 10 - 150 μg/mL. Flow cytometry results showed that FSE induced apoptosis in A549 cells. The proportion of cells in G0/G1-phase increased significantly (p &lt; 0.01), while the proportion of cells in S- and G2/M-phase decreased correspondingly, indicating that the cells were in G0/G1-phase arrest. Cell cycle arrest and apoptosis-inducing effect gradually rose with increase in FSE concentration. With increasing concentrations of FSE, there was also significant increase in the expressions of caspase-3 (p &lt; 0.05), caspase-8 (p &lt; 0.01) and caspase-9 (p &lt; 0.05), but significant decrease in Ki-67 (p &lt; 0.01) and p21 ras protein (p &lt; 0.01).&#x0D; Conclusion: FSE exerts significant inhibitory effect on the proliferation of A549 lung cancer cells. Therefore, the plant can potentially be developed for the treatment of lung cancer.

Evaluation of Liposomal and Microbubbles Mediated Delivery of Doxorubicin in Two-Dimensional (2D) and Three-Dimensional (3D) Models for Breast Cancer.

Liposomal cancer treatment strategies are useful in removing the side effects that were the main concern in recent years. In this study, we prepared microbubble (MBs) conjugated with DOX-loaded liposomes (DOX-loaded MBs) and investigated their effectiveness in in vitro breast cancer cells in two dimensions (2D) and three dimensions (3D). With this aim, breast cancer cells with different features (4T1, MDA-MB231, MCF-7) were growth in 2D and 3D dimensions. The cytotoxic and cell death effects under different conditions, durations and doses were evaluated with WST-1, trypan-blue, colony counts. Apoptotic effects were investigated with flow cytometric Annexin-V-PI and immunohistochemical (Ki-67, caspase 3, 8, 9) methods. After free DOX and LipoDOX were applied, the proliferation index of three cell lines reduced. Intrinsic and extrinsic apoptotic pathways were activated in both 2D and 3D models. However, this effect was observed at lower levels in the 3D model due to the difficulty of diffusion of DOX into the spheroids. Additionally, the suitability of the 3D model for breast cancer cells was supported by formation of ductus-like structures and spheroids. Cell deaths were not observed significantly with the DOX-loaded microbubbles due to rising of MBs to the surface and not reaching spheroids held in matrigel of 3D model. DOX and LipoDOX showed anti-proliferative and apoptosis-inducing effects in breast cancer cells. However, these effects indicated variability depending on the cell lines and 2D or 3D model types.

Imatinib Induces Apoptosis of NK/T Cell Neoplasms by Inhibiting Hematopoietic Cell Kinase and NQO2

▪[Background] Natural killer and T (NK/T)-cell malignancies are uncommon in Western countries but relatively frequent in Asia. Although regimens that include L-asparaginase have improved outcomes, these disorders, particularly aggressive NK-cell leukemias/lymphomas, have poor prognoses. No molecular-targeting drugs have been developed so far. Current reports have provided evidence that a subset of NK cell malignancies is of myeloid origin. Some tyrosine kinase inhibitors (TKIs) that target multiple kinases, including LcK involved in the growth of myeloid malignancies, may therefore show inhibitory effects against NK/T cell neoplasms where LcK members are highly expressed. We investigated the potential anti-tumor activities of multitarget TKIs against malignant NK/T cell neoplasms. [Methods and Results] Three TKIs (imatinib, nilotinib, and dasatinib) were tested for their apoptosis-inducing activities against various NK cell lines including KHYG1, NK92, and NKL after 24-h incubation. Therapeutic doses of imatinib (7.5-27 µM) induced variable annexinV(+) cells in 5 out of 7 NK lines, ranging from 27.1%±6.9% to 87.8%±3.3% (Figure 1), while the apoptosis-inducing effects of therapeutic doses of nilobinib and dasatinib were marginal. Membrane array screenings, which can quantitate 43 phosphorylated proteins, showed that hematopoietic cell kinase (HcK) was constitutively active in KHYG1 cells, and treatment with imatinib inhibited pHcK by 80%. Other known imatinib-targets, including PDGFR, LcK, c-kit, and BCR/ABL, were undetectable in the NK cell lines tested. Further immunoblot analyses using anti-pHcK antibodies confirmed pHcK inhibition in imatinib-treated KHYG1 and NKL (Figure 2). Immunohistochemistry studies with anti-pHcK antibodies revealed two of eight extranodal NK/T-cell lymphomas, nasal type, to be strongly positive for pHcK (Figure 3). In addition, immunohistochemistry studies revealed that NAD(P)H dehydrogenase, quinone 2 (NQO2), a lesser-known imatinib target, was highly expressed in tumor tissues of all seven NK/T-cell lymphoma patients examined, including two pHcK(+) patients, and in vitro studies showed 50% NQO2 protein downregulation and 80% inhibition of its enzymatic activity in imatinib-treated cells. Notably, imatinib enhanced the pro-apoptotic and anti-proliferative activities of L-asparaginase against malignant NK and T cells (CI 0.77 and 0.86 respectively). [Conclusions] Imatinib induces the apoptosis of a subset of NK/T cell malignancies by inhibiting pHcK and NQO2. Imatinib and HcK inhibitors may have therapeutic potential against NK/T cell malignancies either alone or in combination with L-asparaginase. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Anti-proliferation effect of Scutellaria barbata D. Don extract on lung cancer

Purpose: To investigate the effect of Scutellaria barbata D. Don extract (SBDE) on apoptosis and proliferation in A549 human lung cancer cells.&#x0D; Methods: Inverted microscope was used to observe morphological changes in A549 cells after exposure to SBDE. Trypan blue staining of living cells was applied to construct cell growth curve after treatment with varying concentrations of SBDE. The influence of SBDE on cell proliferation, apoptosis and cell cycle was determined by MTT assay while protein expressions of key apoptosis-related enzymes were evaluated by immuno-cytochemical method.&#x0D; Results: SBDE inhibited the growth of A549 lung cancer cells at a concentration range of 20 - 160 μg/mL. Flow cytometry showed that SBDE induced apoptosis in the A549 cells. The proportion of cells in G0/G1-phase increased significantly (p &lt; 0.01), while the proportion of cells in S-phase and G2/Mphase decreased correspondingly, indicating that the cells were in G0/G1-phase arrest. Cell cycle arrest and apoptosis-inducing effect gradually increased with increase in SBDE concentration. With increasing concentrations of SBDE, there were significant increases in the expressions of caspase-3 (p &lt; 0.05), caspase-8 (p &lt; 0.01) and caspase-9 (p &lt; 0.05), and significant decreases in Ki-67 (p &lt; 0.01) and p21 ras protein (p &lt; 0.01).&#x0D; Conclusion: SBDE exerts significant inhibitory effect on the proliferation of A549 lung cancer cells, which can be developed for the treatment of lung cancer patients.

Broad-Spectrum Anticancer Activity and Pharmacokinetic Properties of a Prenyloxy-Substituted Indeno[1,2-b]indole Derivative, Discovered as CK2 Inhibitor.

Protein kinase CK2 is involved in regulating cellular processes, such as cell cycle, proliferation, migration, and apoptosis, making it an attractive anticancer target. We previously described a prenyloxy-substituted indeno[1,2-b]indole (5-isopropyl-4-(3-methylbut-2-enyloxy)-5,6,7,8-tetrahydroindeno[1,2-b]indole-9,10-dione (4p)) as a very potent inhibitor of CK2 holoenzyme (IC50 = 25 nM). Here, we report the broad-spectrum anticancer activity of 4p and provide substantial progress on its pharmacokinetic properties. Using a cell-based CK2 activity assay and live-cell imaging of cultured A431, A549, and LNCaP cancer cell lines, cellular CK2 target engagement was shown as well as strong antiproliferative, anti-migratory and apoptosis-inducing effects of 4p. Furthermore, evidence was found for the ability of 4p to disrupt A549 spheroid cohesion. A series of LC-MS/MS experiments revealed high and rapid cellular uptake (intracellular concentration is approximately 5 µM after 1 h incubation) and low metabolic stability of 4p. These results point to the value of 4p as a potent CK2 inhibitor with promising anticancer activities and should trigger future medicinal chemistry efforts to improve the drug-like properties of this compound.

β-Thujaplicin Enhances TRAIL-Induced Apoptosis via the Dual Effects of XIAP Inhibition and Degradation in NCI-H460 Human Lung Cancer Cells.

Background: β-thujaplicin, a natural tropolone derivative, has anticancer effects on various cancer cells via apoptosis. However, the apoptosis regulatory proteins involved in this process have yet to be revealed. Methods: Trypan blue staining, a WST-8 assay, and a caspase-3/7 activity assay were used to investigate whether β-thujaplicin sensitizes cancer cells to TNF-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis. Additionally, western blotting was performed to clarify the effects of β-thujaplicin on X-linked inhibitor of apoptosis protein (XIAP) in NCI-H460 cells and a fluorescence polarization binding assay was used to evaluate the binding-inhibitory activity of β-thujaplicin against XIAP-BIR3. Results: β- and γ-thujaplicins decreased the viability of NCI-H460 cells in a dose-dependent manner; they also sensitized the cells to TRAIL-induced cell growth inhibition and apoptosis. β-thujaplicin significantly potentiated the apoptosis induction effect of TRAIL on NCI-H460 cells, which was accompanied by enhanced caspase-3/7 activity. Interestingly, β-thujaplicin treatment in NCI-H460 cells decreased XIAP levels. Furthermore, β-thujaplicin was able to bind XIAP-BIR3 at the Smac binding site. Conclusions: These findings indicate that β-thujaplicin could enhance TRAIL-induced apoptosis in NCI-H460 cells via XIAP inhibition and degradation. Thus, the tropolone scaffold may be useful for designing novel nonpeptidic small-molecule inhibitors of XIAP and developing new types of anticancer drugs.

Cyclovirobuxine D Induced-Mitophagy through the p65/BNIP3/LC3 Axis Potentiates Its Apoptosis-Inducing Effects in Lung Cancer Cells.

Mitophagy plays a pro-survival or pro-death role that is cellular-context- and stress-condition-dependent. In this study, we revealed that cyclovirobuxine D (CVB-D), a natural compound derived from Buxus microphylla, was able to provoke mitophagy in lung cancer cells. CVB-D-induced mitophagy potentiates apoptosis by promoting mitochondrial dysfunction. Mechanistically, CVB-D initiates mitophagy by enhancing the expression of the mitophagy receptor BNIP3 and strengthening its interaction with LC3 to provoke mitophagy. Our results further showed that p65, a transcriptional suppressor of BNIP3, is downregulated upon CVB-D treatment. The ectopic expression of p65 inhibits BNIP3 expression, while its knockdown significantly abolishes its transcriptional repression on BNIP3 upon CVB-D treatment. Importantly, nude mice bearing subcutaneous xenograft tumors presented retarded growth upon CVB-D treatment. Overall, we demonstrated that CVB-D treatment can provoke mitophagy and further revealed that the p65/BNIP3/LC3 axis is one potential mechanism involved in CVB-D-induced mitophagy in lung cancer cells, thus providing an effective antitumor therapeutic strategy for the treatment of lung cancer patients

Maclurin exerts anti-cancer effects in human osteosarcoma cells via prooxidative activity and modulations of PARP, p38, and ERK signaling.

Maclurin [(3,4-dihydroxyphenyl)-(2,4,6-trihydroxyphenyl) methanone] is a natural compound that can be extracted from white mulberry(Morus alba) and purple mangosteen(Garcinia mangostana). Maclurin is known for its dual-sided effect on reactive oxygen species (ROS). Osteosarcoma is a primary malignant tumor of the bone and is one of the most aggressive cancers. It is common especially in children and young adults and can progress into highly metastatic cancer. In this study, we investigated the anti-cancer effects of maclurin on U2OS human osteosarcoma cells. The results indicated that maclurin exerts prooxidative effects and induces apoptosis via capase-3-independent PARP regulation in U2OS human osteosarcoma cells. Maclurin also inhibits the migration of U2OS human osteosarcoma cells. Maclurin modulates two of the three major mitogen-activated protein kinases that are closely linked with cancer metastasis; that is, it activates p38 and inactivates Extracellular signal-regulated kinase. The apoptosis-inducing effects of maclurin on U2OS osteosarcoma cells were diminished by additional treatment with antioxidant N-acetyl cysteine (NAC), but the migration-inhibiting effect was not affected by NAC treatment. This further suggested the only apoptosis-inducing effect of maclurin may be strongly related to its prooxidative activity. Taken together, these results suggested that maclurin may be a strong candidate molecule as an anti-osteosarcoma agent.

Pinus kesiya Royle ex Gordon induces apoptotic cell death in hepatocellular carcinoma HepG2 cell via intrinsic pathway by PARP and Topoisomerase I suppression

Pinus kesiya Royle ex Gordon (PK), widely found in Southeast Asia, has been traditionally used for the treatment of several illnesses. Our previous studies showed that PK was highly cytotoxicity against liver cancer cells. The detailed mechanism of anticancer action of 50% hydro-ethanolic extract of PK’s twig was, therefore, investigated in hepatocellular carcinoma HepG2 cells. Cytotoxicity of PK was determined by using NR assay, followed by determination of the mode of cell death by flow cytometry. The apoptosis-inducing effect was determined based on caspases activity, mitochondria membrane potential change, and expression of proteins related to apoptosis by western blot. The biomolecular alteration in the PK-treated HepG2 cells was investigated by FTIR microspectroscopy. Inhibition of topoisomerase I enzyme was determined by using DNA relaxation assay. Results showed that PK displayed high selective cytotoxicity and induced apoptosis against HepG2. FTIR microspectroscopy indicated that PK altered major biomolecules in HepG2 different from melphalan (a positive control), indicating a different mechanism of anticancer action. PK induced apoptotic cell death through the intrinsic pathway by increasing caspases 9 and 3/7 activity, increasing Bax, and decreasing Bcl-2 expression leading to mitochondrial membrane potential changes. PK also inhibited Top I and PARP activity that triggered an intrinsic apoptotic pathway. The phytochemical test presented terpenoids (i.e., α-pinene confirmed by GC-MS), alkaloids, steroids, xanthone, reducing sugar, and saponin. α-Pinene exhibited low cytotoxicity against HepG2, therefore, several terpene derivatives may work synergistically for inducing apoptosis. Our data demonstrated that PK has the potential for further study with chemotherapeutic purposes.

CDK7 blockade suppresses super-enhancer-associated oncogenes in bladder cancer.

Transcriptional addiction plays a pivotal role in maintaining the hallmarks of cancer cells. Thus, targeting super-enhancers (SEs), which modulate the transcriptional activity of oncogenes, has become an attractive strategy for cancer therapy. As yet, however, the molecular mechanisms of this process in bladder cancer (BC) remain to be elucidated. Here, we aimed to provide detailed information regarding the SE landscape in BC and to investigate new potential pharmaceutical targets for BC therapy. We employed THZ1 as a potent and specific CDK7 inhibitor. In vitro and in vivo studies were carried out to investigate the anticancer and apoptosis-inducing effects of THZ1 on BC cells. Whole-transcriptome sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) were performed to investigate the mechanism and function of SE-linked oncogenic transcription in BC cells. We found that THZ1 serves as an effective and potent inhibitor with suppressive activity against BC cells. An integrative analysis of THZ1-sensitive and SE-associated oncogenes yielded potential new pharmaceutical targets, including DDIT4, B4GALT5, PSRC1 and MED22. Combination treatment with THZ1 and the DDIT4 inhibitor rapamycin effectively suppressed BC cell growth. In addition, we found that THZ1 and rapamycin sensitized BC cells to conventional chemotherapy. Our data indicate that exploring BC gene regulatory mechanisms associated with SEs through integrating RNA-seq and ChIP-seq data improves our understanding of BC biology and provides a basis for innovative therapies.

Altered Long Non-coding RNAs Expression and Cytotoxic and Anti-proliferative Activity of Dendrosomal Nano-curcumin in Ovarian Cancer Cells

Curcumin is a polyphenol derived from the rhizome of turmeric, which has significant chemoprevention and chemotherapeutic effects. Long non-coding RNAs (lncRNAs) are longer than 200 nucleotides and play a crucial role in various biological processes. Since aberrant lncRNAs expression affects the development and progression of tumors, the present study was conducted to determine the cytotoxic potential of dendrosomal nano-curcumin (DNC) against ovarian cancer cell proliferation and apoptosis and altered expression of some lncRNAs involved in ovarian cancer. In vitro cytotoxic activity of DNC against OVCAR3 and SKOV3 human ovarian cancer cells and HFSF-PI3 human fibroblasts cells as a control was evaluated by MTT assay. Real-time PCR and western blot methods were used to determine the apoptosis-inducing effect of DNC through analyzing apoptosis marker genes and protein expression, respectively. We also checked the ovarian cancer cells apoptosis induced with change in expression levels of some lncRNAs. Our results revealed that DNC significantly decreased viability of ovarian cancer cells as compared to Cur. Moreover, DNC reduced the relative expression levels of HOTAIR and H19 and increased the expression of MEG3. Bcl2 expression was down-regulated in mRNA and protein level after DNC treatment, whereas knockdown of MEG3 reduced it. Our findings discovered the effect of DNC on growth inhibition as well as modulation of some lncRNAs in ovarian cancer cells. We also observed that MEG3 down-regulation can suppress the effect of DNC on down-regulation of Bcl-2 as an anti-apoptotic protein. This may suggest the role of MEG3 in DNC-induced apoptosis.

Hematoxylin binds to mutant calreticulin and disrupts its abnormal interaction with thrombopoietin receptor

Somatic mutations of calreticulin (CALR) have been identified as a main disease driver of myeloproliferative neoplasms, suggesting that development of drugs targeting mutant CALR is of great significance. Site-directed mutagenesis in the N-glycan binding domain (GBD) abolishes the ability of mutant CALR to oncogenically activate the thrombopoietin receptor (MPL). We therefore hypothesized that a small molecule targeting the GBD might inhibit the oncogenicity of the mutant CALR. Using an in silico molecular docking study, we identified candidate binders to the GBD of CALR. Further experimental validation of the hits identified a group of catechols inducing a selective growth inhibitory effect on cells that depend on oncogenic CALR for survival and proliferation. Apoptosis-inducing effects by the compound were significantly higher in the CALR-mutated cells than in CALR wild-type cells. Additionally, knockout or C-terminal truncation of CALR eliminated drug hypersensitivity in CALR-mutated cells. We experimentally confirmed the direct binding of the selected compound to CALR, disruption of the mutant CALR-MPL interaction, inhibition of the JAK2-STAT5 pathway, and reduction at the intracellular level of mutant CALR upon drug treatment. Our data indicate that small molecules targeting the GBD of CALR can selectively kill CALR-mutated cells by disrupting the CALR-MPL interaction and inhibiting oncogenic signaling.