Expression Of Anti-apoptotic Proteins Research Articles

Mitochondrial Localization of Antioxidant Nanodrug Suppresses Ocular Inflammation by Alleviating Oxidative Stress on Cells.

External environments (e.g., pollutants, irritants, ultraviolet radiation, etc) probably activate oxidative stress on the ocular surface, further leading to inflammatory responses and cellular apoptosis. For treating ophthalmic diseases, one of the strategies is to regulate oxidative stress through antioxidants. Here we conjugate a polyphenolic antioxidant drug (i.e., caffeic acid) with a small peptide of protein tag to generate a peptide-drug conjugate as a nanodrug. With a self-assembled ability to form nanoparticles, the nanodrug mainly enters human corneal epithelial cells (HCEC) by caveolin-mediated endocytosis, succeeds lysosomal escape, and achieves mitochondrial localization of caffeic acid. Revealed by free radical scavenging experiments, the nanodrug shows considerable antioxidant capacities. In mouse leukemia cells of monocyte macrophage (RAW 264.7) induced by lipopolysaccharide (LPS), the nanodrug inhibits various pro-inflammatory cytokines (e.g., NO, IL-6, and TNF-α) by up-regulating the expression of anti-apoptotic protein (e.g., Bcl-2). As an investigation of alleviating oxidative stress by the mitochondrial localization of antioxidant, this work may establish an experimental foundation for the regulation of cellular redox balance, as well as provide different insights for the clinical development of antioxidant drug delivery systems in the treatment of ocular disease.

Chitosan oligosaccharide alleviates DON-induced liver injury via suppressing ferroptosis in mice.

Chitosan oligosaccharide (COS), a water-soluble derivative of chitin, has been recognized for its diverse biological properties. Deoxynivalenol (DON) is a prevalent mycotoxin, causing extreme liver damage. However, the mechanism whereby COS alleviates DON-induced liver injury remains unclear. In the present study, C57BL/6 mice were randomly divided into four groups: control (CON), DON (1.0 mg/d/kg BW DON), COS (200 mg/d/kg BW COS), and COS+DON (200 mg/d/kg BW COS + 1.0 mg/d/kg BW DON), with a period of 28 days. The results indicated that COS effectively reversed DON-induced weight loss, elevated liver index, and liver hemorrhage and swelling in mice. Moreover, COS significantly reduced liver reactive oxygen species (ROS) levels, malondialdehyde (MDA) content, and lactate dehydrogenase (LDH) release in DON-exposed mice, while restoring the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC). Further investigations revealed that COS modulated the expressions of pro-inflammatory cytokines and anti-apoptotic proteins through stimulation of the Nrf2/HO-1 signaling pathway and suppression of the NF-κB signaling pathway. Additionally, COS inhibited ferroptosis by modulating the SLC7A11/GSH/GPX4 pathway and the expression of FTH1 and FLC proteins, thereby reducing lipid peroxidation accumulation and iron overload. In summary, this research showed that COS mitigated DON-induced liver injury in mice by alleviating DON-induced oxidative stress, inflammation, apoptosis, and ferroptosis via modulating the Nrf2/HO-1/NF-κB and GPX4 signaling pathways. These results offer a theoretical basis for the development and application of COS as a novel liver protectant and propose innovative therapeutic strategies for combating DON-induced liver damage.

Bullatine A suppresses glioma cell growth by targeting SIRT6.

Gliomas are the most common primary tumors of the nervous system, which is generally treated using adjuvant chemotherapy following surgical resection. However, patient survival time is still short, and there is currently no successful treatment for highly malignant gliomas. Bullatine A (BLA) is a diterpenoid alkaloid of the genus Aconitum which antirheumatic and anti-inflammatory pharmacological properties. The effects of BLA on gliomas have not yet been elucidated. In this study, we investigated the effects of BLA on human brain malignant glioblastoma cells. Our results showed that BLA inhibited the proliferation of U87MG and U251cells in a dose-dependent manner and decreased their survival rate. BLA dose-dependently induced apoptosis in U87MG cells, upregulated the expression of cleaved caspase-9, cleaved caspase-3 pro-apoptotic protein, and Bax protein, and downregulated the expression of Bcl-2 anti-apoptotic protein. Moreover, BLA dose-dependently induced U87MG and U251cell cycle arrest in the G2/M phase, and downregulated the expression of p-ERK and Myc proteins. Further, BLA significantly inhibited the acetylation of histones H3K9 and H3K56, and upregulated the expression of the protein deacetylase SIRT6. Mechanistic studies revealed that the effect of BLA on inducing apoptosis and inhibiting the proliferation of glioma cells was blocked by SIRT6 knockout. In summary, our study indicated that BLA is a potential therapeutic agent for glioma that targets SIRT6 to inhibit glioma cell proliferation and induce apoptosis.

Discovery of novel JQ1 derivatives as dual ferroptosis and apoptosis inducers for the treatment of triple-negative breast cancer.

The activation of ferroptosis in refractory cancers may enhances their sensitivity to apoptosis-based chemotherapy, resulting in a synergistic effect via combination therapy. To enhance the anticancer effect of JQ1, a known BRD4 inhibitor with a significant antiproliferative effect on triple-negative breast cancer (TNBC), various new JQ1 derivatives as dual ferroptosis and apoptosis inducers were designed and synthesized. Among them, compound BG11 revealed a remarkable inhibitory activity against TNBC cells and obviously suppressed BRD4 and GPX4 expression and activities. Further studies suggested that BG11 induced cell ferroptosis through promoting Fe2+ and intracellular lipid peroxide deposition. In addition, BG11 could induce apoptosis through increasing Bax (apoptotic protein) expression and decreasing Bcl-2 (anti-apoptotic protein) expression within MDA-MB-231cells. Surprisingly, BG11 significantly inhibited tumor proliferation in the MDA-MB-231 xenograft model without obvious toxicity. Based on the above findings, BG11 may be the candidate dual ferroptosis and apoptosis inducers for treating TNBC.

Smilaxchina L. polyphenols inhibit LPS-induced macrophage M1 polarization to alleviate inflammation through NF-κB signaling pathway in vitro and in vivo.

As an important component of the cell wall of Gram-negative bacteria, lipopolysaccharide (LPS) is an important inducer of inflammation in humans. Smilax china L. is known for its diverse bioactive functions, particularly its anti-inflammatory effects. This study aimed to investigate the bioactive function of Smilax china L. polyphenols (SCLP) on LPS-induced inflammation. Inflammation in RAW264.7 macrophages and mice were induced using LPS. The cytotoxicity of SCLP was investigated by MTT assay. Inflammatory factors were detected by ELISA and RT-PCR. The expression of NF-κB pathway-related proteins was analyzed by Western Blotting. The results demonstrated that SCLP significantly reduced the levels of pro-inflammatory factors (TNF-α, IL-1β, and IL-6) and inhibited M1 polarization of macrophages in both RAW264.7 macrophages and mice (p<0.05). Western Blotting analysis revealed that the levels of NF-κB signaling pathway-associated proteins (p-p65, p-IKB, p-IKK) were significantly reduced (p<0.05). Notably, SCLP significantly downregulated the expression of pro-apoptotic proteins, while upregulating the expression of anti-apoptotic proteins in RAW264.7 macrophages (p<0.05). Additionally, the levels of antioxidant enzymes were enhanced in mice, suggesting a potential reduction in the inflammatory response. These findings indicated that SCLP might inhibit LPS-induced M1 polarization through the NF-κB signaling pathway, thereby reducing inflammation. Consequently, SCLP might serve as a promising bioactive substance for preventing inflammation-related injury.

ML385, a selective inhibitor of Nrf2, demonstrates efficacy in the treatment of adult T-cell leukemia

Nrf2 plays a critical role in regulating cytoprotective transcriptional responses and glucose metabolism while also preventing inflammation-induced carcinogenesis. However, Nrf2 can paradoxically promote carcinogenesis. Here, we aimed to elucidate the role of Nrf2 in ATL associated with HTLV-1. HTLV-1-infected T-cell lines exhibited nuclear accumulation of Nrf2. Nrf2 knockdown along with the inhibition of its activity using ML385, decreased cell proliferation and survival. Furthermore, ML385-induced G1 arrest by enhancing γH2AX and p53 expression while downregulating CDK4/6, cyclin D2/E, and c-Myc. Additionally, ML385 triggered caspase-mediated apoptosis by downregulating the expression of anti-apoptotic proteins while upregulating pro-apoptotic proteins. The compound also induced necroptosis, promoted JNK phosphorylation, and inhibited the NF-κB, AP-1, and STAT3/5 signaling. Moreover, ML385 was found to reduce the expression of LDHA, glucose uptake, and the levels of lactate derived from glycolysis. Overall, these results suggest that Nrf2 functions as an oncogene in ATL and may represent a promising therapeutic target.

Garcinol, a natural benzophenone overcomes radio and chemoresistance by targeting PI3K-PKB/AKT pathway in rhabdomyosarcoma cells

AKT signaling pathway is a critical intracellular signaling pathway involved in regulating various cellular processes, including cell proliferation, cell survival, cell migration and metabolism. Cancer cells have a propensity to alter the expression of pro- and anti-angiogenic signals to activate the angiogenic switch. Aberrant overexpression of AKT pathway is associated with various types of cancers and enables the migration and invasion of tumor cells to neighboring tissues. Garcinol, a natural benzophenone, displays potent anticancer potential against various cancer cells, but its mechanism of action in inducing apoptosis remains unclear. The present study aimed to unveil the mechanism of action of garcinol in inducing apoptosis in rhabdomyosarcoma cells. Garcinol inhibited cell proliferation, reduced the viability of cancer cells, diminished colony formation, and mitigated the migratory capabilities of Rh cells. In vitro analysis of garcinol unveiled potent anticancer and anti-metastatic activity, with an IC50 ranging from 5.32 to 16.28 against Rh cell lines, as demonstrated by MTT assay. Garcinol actuates apoptosis by triggering mitotic arrest (G2/M phase arrest) in Rh cell lines and alters the expression of pro-apoptotic and anti-apoptotic proteins to trigger efficient apoptosis in cancer cells.

Design, synthesis, and antiproliferative activity evaluation of novel α-mangostin derivatives by ROS/MAPK signaling pathway

Novel hydroxamic acid and 3,6-amide modified α-mangostin derivatives were synthesized and evaluated their antiproliferative activities against KYSE 30 (esophageal cancer), HCT 116 (colon cancer), and HGC 27 (gastric cancer) cell lines. Most of the new derivatives displayed stronger anti-proliferative activities compared to α-mangostin. Among all the derivatives, compound 4a exhibited the most potent activity, with IC50 values of 0.57 ± 0.29 μM, 3.27 ± 0.16 μM, and 2.28 ± 1.02 μM against KYSE 30, HCT 116, and HGC 27 cells, respectively. Subsequent mechanism studies revealed that compound 4a inhibited cancer cells proliferation and colonies formation in a concentration-dependent manner. Additionally, compound 4a caused cell cycle arrest in a p53 dependent manner and induced apoptosis in p53 independent way. Meanwhile, 4a suppressed cell cycle related proteins (Cyclin D1 and cyclin B1) expression, increased pro-apoptotic proteins (cleaved PARP, cleaved caspase-7, and cleaved caspase-9) and decreased anti-apoptotic proteins (Bcl-2) expression. Moreover, 4a increased reactive oxygen species (ROS) levels in KYSE 30 cells and upregulated the expression of proteins related to the ROS related MAPK signaling pathway (p-ERK, p-p38, and p-JNK). These findings suggest that compound 4a holds promising potential as an antiproliferative agent by targeting MAPK signaling pathway to inhibit cell cycle progress, induce apoptosis and produce ROS in cancers.

CTNI-61. PRELIMINARY RESULTS OF A PHASE I/II TRIAL OF SELINEXOR AND TEMOZOLOMIDE IN RECURRENT GLIOBLASTOMA

Abstract BACKGROUND Selinexor (SEL) is a first-in-class XPO1 inhibitor with potent antitumor activity through the nuclear retention and reactivation of tumor suppressor proteins, reduced translation of oncogenes, and decreased expression of anti-apoptotic proteins. As a single agent, SEL has demonstrated brain penetration and clinically relevant responses in glioblastoma. We seek to enhance the effect and benefit of SEL by priming with temozolomide (TMZ). METHODS Through an NCI Project Team, we developed a multi-institutional phase I/II clinical trial which opened across the NCI Experimental Therapeutics Clinical Trial Network. Eligibility included histologically confirmed 1st recurrent MGMT promoter methylated glioblastoma. We evaluated safety, tolerability, and dose finding with an IQ 3 + 3 design of TMZ 150mg/m2 on days 1-5 of a 28-day cycle plus SEL on days 8 and 15. RESULTS From October 2022 to March 2024, we enrolled 12 patients (n=11 evaluable) into phase I of the study. Patient characteristics included 82% male, median age 62 (range 50-77), 64% white. The median number of cycles administered was 6 (range 1-12). Two dose levels (SEL 60mg and SEL 80mg) were evaluated. No dose-limiting toxicities (DLTs) were observed. The most common treatment related adverse events were nausea (58%), decreased platelet count (50%), fatigue (50%), constipation (42%), vomiting (25%), decreased lymphocyte count (25%), and decreased neutrophil count (25%). Grade 3+ treatment related adverse events included nausea, grade 3 (n=1); decreased lymphocyte count, grades 3-4 (n=2); and anorexia, grade 3 (n=1). Phase I has completed enrollment and the recommended phase II dose will be SEL 80mg. Additional results will be presented. CONCLUSION Results suggest that a sequential dosing regimen of SEL+TMZ is feasible, well-tolerated, and may minimize the cumulative toxicity of SEL. The phase II randomized portion of this trial is enrolling nationwide and will investigate efficacy and candidate molecular signatures of vulnerability (NCI #10505, NCT05432804).

Combined Targeting of SYK and BCL-2 in Acute Myeloid Leukaemia and Mechanism Study

Acute myeloid leukemia (AML) is a highly heterogeneous haematological malignancy and is the most common acute leukaemia in adults, with a median age of onset of 68 years and a 5-year survival rate of only 30%. Only 60-80% of adult AML patients can achieve complete remission with the first induction chemotherapy, and 50-80% of AML patients ultimately relapse with the “3+7” first-line treatment regimen based on anthracycline and cytarabine (Ara-C), so there is an urgent need to find safer and more effective targeted therapies. There is an urgent need to find safer and more effective targeted therapies. Venetoclax (ABT-199) is an FDA-approved oral selective Bcl-2 inhibitor with limited anti-leukaemia activity as a single agent, and mechanisms of resistance include up-regulation of anti-apoptotic proteins, compensation of multiple metabolic pathways such as OXPHOS glycolysis, and mutations in BCL-2 and BAX.In 2018, the FDA approved the use of Venetoclax in combination with demethylating drugs (azacitidine ( AZA) or low-dose azacitidine AraC (LDAC)) in combination for newly diagnosed AML in older adults aged 75 years and older, and while this combination greatly improves remission rates, most patients relapse within 18 months. It has been suggested that direct or indirect inhibition of oxidative phosphorylation (OXPHOS) may be able to circumvent adaptive resistance of AML to Venetoclax+ AraC combination therapy. Therefore, an ideal candidate therapy in combination with Venetoclax should be able to antagonise its resistance mechanisms from metabolic pathways and down-regulation of MCL-1. Purpose To investigate the efficacy and related mechanisms of co-targeting SYK with BCL-2 for the treatment of acute myeloid leukaemia (AML). Experimental Design AML cell lines and primary patient samples were collected for cell proliferation assays. Firstly, cell activity was detected by MTS colorimetric assay, by Compusyn calculation software, the joint index CI value was calculated, apoptosis was detected by flow cytometry, mitochondrial membrane potential was altered, gene expression was detected by transcriptome sequencing, and the signalling pathways were analysed, followed by apoptosis-related proteins, BCL-2-related proteins by Western Blot, and glucose was detected by metabolism kit. sugar consumption,ATP and lactate content. The in vivo efficacy of the combination of Venetoclax and Entospletinib was assessed using an MV4-11-derived xenograft mouse model. Results SYK is highly expressed in AML and is associated with poor prognosis. ...The combination of Venetoclax and Entospletinib in AML cell lines and AML primary patient cells had significant synergistic growth inhibitory and apoptosis-inducing effects...Venetoclax in combination with Entospletinib down-regulated the expression of anti-apoptotic proteins, MCL-1, and BCL-XL, and down-regulated phosphorylated ERK expression, thereby affecting MCL1 post-translational protein stability rather than transcriptional levels.... Venetoclax in combination with Entospletinib downregulated the expression of anti-apoptotic proteins MCL-1, BCL-XL and phosphorylated ERK, thereby affecting the post-translational protein stability of MCL1, but not the transcriptional level. Conclusions The combination of Venetoclax and Entospletinib had significant synergistic killing effects in both AML cell lines and AML primary patient cells.Venetoclax in combination with Entospletinib exerts synergistic therapeutic effects on AML through down-regulation of ERK phosphorylation affecting the expression of anti-apoptotic proteins MCL-1, BCL-XL and reprogramming of glucose metabolism.

Protective Role of Sphingosine-1-Phosphate During Radiation-Induced Testicular Injury.

The impact of ionizing radiation on the male reproductive system is gaining increasing attention, particularly when it comes to testicular damage, which may result in decreased sperm quality and hormonal imbalances. Finding effective protective measures to mitigate testicular damage caused by radiation has become a focal point in the biomedical field. S1P, an essential biological signaling molecule, has garnered significant interest due to its multiple roles in regulating cellular functions and its protective effects against radiation-induced testicular injury. S1P not only effectively reduces the generation of ROS induced by radiation but also alleviates oxidative stress by enhancing the activity of antioxidant enzymes. Furthermore, S1P inhibits radiation-induced cell apoptosis by regulating the expression of anti-apoptotic and pro-apoptotic proteins. Additionally, S1P alleviates radiation-induced inflammation by inhibiting the production of inflammatory factors, thereby further protecting testicular tissue. In summary, S1P effectively reduces radiation-induced testicular damage through multiple mechanisms, offering a promising therapeutic approach to safeguard male reproductive health. Future research should explore the specific mechanisms of action and clinical application potential of S1P, aiming to contribute significantly to the prevention and treatment of radiation damage.

Noradrenaline Protects Human Microglial Cells (HMC3) Against Apoptosis and DNA Damage Induced by LPS and Aβ1-42 Aggregates In Vitro.

Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder, characterized by the accumulation of amyloid-beta (Aβ) plaques and neuroinflammation. This study investigates the protective effects of noradrenaline (NA) on human microglial cells exposed to lipopolysaccharides (LPS) and Aβ aggregates-major contributors to inflammation and cellular damage in AD. The reduced Aβ aggregation in the HMC3 human microglial cells co-treated with Aβ and NA was confirmed by thioflavin T (ThT) assay, fluorescent ThT staining, and immunocytochemistry (ICC). The significantly increased viability of HMC3 cells after 48 h of incubation with NA at 50 µM, 25 µM, and 10 µM, exposed to IC50 LPS and IC50 Aβ, was confirmed by XTT and LDH assays. Moreover, we found that NA treatment at 25 μM and 50 μM concentrations in HMC3 cells exposed to IC50 LPS or IC50 Aβ results in an increased proliferation of HMC3 cells, their return to normal morphology, decreased levels of DNA damage, reduced caspase-3 activity, decreased expression of pro-apoptotic DDIT3 and BAX, and increased expression of anti-apoptotic BCL-2 genes and proteins, leading to enhanced cell survival, when compared to that of the HMC3 cells treated only with IC50 LPS or IC50 Aβ. Furthermore, we showed that NA induces the degradation of both extracellular and intracellular Aβ deposits and downregulates hypoxia-inducible factor 1α (HIF-1α), which is linked to impaired Aβ clearance and AD progression. These findings indicate that NA holds promise as a therapeutic target to address microglial dysfunction and potentially slow the progression of AD. Its neuroprotective effects, particularly in reducing inflammation and regulating microglial activity, warrant further investigation into its broader role in mitigating neuroinflammation and preserving microglial function in AD.

Gochujang suppresses cell survival and changes reactive oxygen species metabolism in colorectal cancer cells.

There is a significant global increase in colorectal cancer (CRC) among young adults. Gochujang, one of the signature Korean traditional fermented foods, contains various bioactive compounds and has multiple health-beneficial effects, including anticancer effects; however, the detailed cellular and molecular mechanisms of its anticancer outcomes are not fully understood. The objective of the present study was to investigate the detailed underlying anticancer mechanisms of Gochujang in CRC cells. Gochujang was extracted with 80% ethanol, and total polyphenol contents (9.9 ± 1.63 mgGAE/g) and total flavonoid contents (0.14 ± 0.07 mgQE/g) of Gochujang extract (GE) were evaluated. GE significantly suppressed cell viability, migration, and colony formation in CRC cells. Also, GE increased the cell cycle arrest-related protein p21 level, whereas it decreased cell cycle progression-associated proteins, such as p-Rb. Moreover, GE markedly elevated the levels of proapoptotic proteins (e.g. Bim and c-PARP), while it downregulated antiapoptotic protein expressions (e.g. Bcl-2 and Bcl-xL). GE also altered the expression of the autophagy-involved proteins. Furthermore, GE strongly reduced the expression of major antioxidant enzymes and increased the reactive oxygen species (ROS) generation in CRC cells, causing an imbalance of ROS metabolism. In conclusion, this study demonstrated that Gochujang exerts anticancer effects in CRC cells by inhibiting cell proliferation, increasing cell death, and interrupting ROS metabolism.

Neuroprotective Potential of Ethoxzolamide Targeting Oxidative Stress and Inflammation in Experimental Models of Intracerebral Hemorrhage.

As antioxidant and anti-inflammatory agents, carbonic anhydrase inhibitors can exert potentially useful therapeutic effects following central nervous system trauma, including intracerebral hemorrhage (ICH). However, the therapeutic efficacy of ethoxyzolamide (ETZ) as a novel carbonic anhydrase inhibitor for ICH has not yet been determined. An autologous blood injection method was used to establish ICH models, which were then used to establish the effects of intraperitoneal injection of ETZ on ICH. Neuronal damage, apoptotic protein expression, oxidative and inflammatory factor content, microglia marker Iba-1 positivity, hepatic and renal pathological changes, and serum concentrations of hepatic and renal function indices were assessed by Nissl staining, western blotting, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, hematoxylin and eosin (HE) staining, and automatic biochemical analysis in brain tissues. The ICH group showed massive hemorrhagic foci; significant increases in brain water content, modified mouse neurological deficit scoring (mNSS) score, pro-apoptotic protein expression, oxidative factors, pro-inflammatory factors, and Iba-1 positivity; and significant reductions in Nissl body size, anti-apoptotic protein expression, and antioxidant factors, all of which were reversed by ETZ in a dose-dependent manner. ETZ has a good biosafety profile with no significant burden on the human liver or kidneys. The Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway was mildly activated in ICH mice, and was further increased after ETZ injection. Molecular docking experiments revealed that ETZ could dock onto the Nrf2-binding domain of keap1. ETZ, as a novel carbonic anhydrase inhibitor, further activated the Keap1/Nrf2 pathway by docking with the Nrf2-binding domain of keap1, thereby exerting antioxidant, anti-inflammatory, anti-apoptotic, and cerebral neuroprotective effects in ICH mice.

Transforming growth factor-beta and Epithelial-mesenchymal transition is considered an attractive pivotal Genetic-Biomarker for Hepatocellular carcinoma with hepatitis C virus-infection patients

Abstract Introduction/Objective Hepatocellular carcinoma (HCC) is the most common cause of cancer worldwide. HCC is induced by hepatitis C-virus (HCV) which represents a major health problem in Egypt. The transforming growth factor- beta (TGF-β) is a potent cytokine that has a multifunctional pro-fibrotic activity, for enhancing liver-inflammation, fibrosis, tumor-progression, and metastasis. TGF-β has a vital role in the regulation of tumorigenesis, by controlling epithelial-mesenchymal-transition (EMT), apoptosis, proliferation, and differentiation process. The dysregulation of TGF-β, Wnt/β-catenin signaling pathways, by HCV-core protein is implicated in the development of HCC. Methods/Case Report The study aims to determine the effect of the HCV-infection in HCC patients with a set of growth-factors, EMT, and anti-apoptotic protein expression levels by using western blotting and immunohistochemistry staining analysis. Results (if a Case Study enter NA) These results observed an insignificant increase in the expression of TGF-β protein for HCC/HCV-infected cells compared to the HCV-uninfected cells, and the expression of the β-catenin protein level is elevated in the nucleus. Also, it showed that Bcl-2 protein is a significant increase in HCV-infected when compared to HCV-free groups. Positive staining for p53 protein was detected in the HCC/HCV-infected, more than in HCC/HCV-uninfected cells. This data indicated that the core-protein of HCV can switch TGF-β from a tumor suppressor to a tumor promoter by suppressing apoptosis of hepatocytes and increasing EMT expression by activation of Wnt/β-catenin signaling pathway. Therefore, HCV-core protein enhances the nuclear accumulation of β-catenin expression protein to activate downstream target genes, which regulate cell-growth and cell-cycle progression. Conclusion It is speculated that HCV may exert an anti-apoptotic effect, and thus enhance tumorigenesis. Also, the accumulation of nuclear Wnt/β-catenin and p53 correlates with TGF-β expression, typically a late event in hepato- carcinogenesis. Taken together, the expression level of TGF-β, Wnt/β-catenin, Bcl-2, and p53 genotype in HCV is considered an indicator of shortened survival in patients, and may be an attractive clinically pivotal genetic biomarker to mediate liver pathogenesis.

Pseudomonas aeruginosa-Mannose Sensitive Hemagglutinin-based Treatment Strategy for Liver Cancer

Background Although it does not represent a threat to healthy humans, Pseudomonas aeruginosa is a type of aerobic, gram-negative bacteria that can cause catastrophic infections in patients with immune system abnormalities and cystic fibrosis. Purpose The current study investigated the effect of Pseudomonas aeruginosa-mannose sensitive hemagglutinin (PA-MSHA) on liver cancer cell viability, colony formation, and invasion potential, and explored the underlying mechanism. Methods Proliferation potential and clonogenic survival of the cells were studied using MTT and colony formation assays, respectively. Transwell and Western blotting assays were used for the assessment of invasive potential and protein expression, respectively. Results The results revealed that exposure to PA-MSHA led to a time-dependent suppression in HepG2 and HEP3B cell proliferation. Incubation with PA-MSHA decreased HepG2 cell proliferation to 92%, 75%, 50%, 35%, and 20%, respectively, after 12, 24, 36, 48, and 72 hours. Similarly, incubation for 12, 24, 36, 48, and 72 hours reduced the proliferation of HEP3B cells to 94%, 79%, 55%, 41%, and 26%, respectively. The rate of clonogenic survival was significantly lower in HepG2 and HEP3B cells on incubation with PA-MSHA. The cell invasion potential was also significantly reduced on incubation with PA-MSHA. In HepG2 cells, incubation with PA-MSHA significantly reduced the expression of Bcl-2 (26 kDa) protein and promoted Bax (21 kDa) level. Following incubation with PA-MSHA, HepG2 cells showed higher cleaved caspase-3 level compared to the control cells. Compared to control cells, the PA-MSHA incubation of HepG2 cells resulted in a prominent reduction in Notch3 protein expression. Conclusion In summary, PA-MSHA treatment prevents liver cancer cell proliferation and targets the survival of clonogenic cells. It reduces invasiveness, targets Notch3 protein expression in liver cancer cells, and increases the level of proapoptotic and suppresses antiapoptotic protein expression. Therefore, PA-MSHA shows encouraging anticancer effects on HepG2 and HEP3B cancer cells, suggesting that it could be developed as a viable treatment option for liver cancer.

A Dual Action Platinum(IV) Complex with Self-assembly Property Inhibits Prostate Cancer through Mitochondrial Stress Pathway.

Platinum(IV) prodrugs are highly promising anticancer agents because they can selectively target tumors and minimize the adverse effects associated with their PtII congeners. In this study, we synthesized dual action PtIV complexes by linking oxoplatin with lithocholic acid. The synthesized compounds, designated as PL-I, PL-II, and PL-III, can spontaneously self-assemble in water, resulting in the formation of spherical shape nanoparticles. Among the developed complexes, PL-III appeared to be the most potent compound against all the tested cancer cell lines, with 10 fold higher cytotoxicity compared to cisplatin in PC3 cells. The complex arrests the cell cycle in the S and G2 phases and induces DNA damage. Additional mechanistic investigations demonstrate that PL-III predominantly localizes within the mitochondria and cytoplasm. Consequently, PL-III disrupts mitochondrial membrane potential, increases ROS production, and perturbs mitochondrial bioenergetics in PC3 cells. The complex induces apoptosis through the mitochondrial pathway by upregulating pro-apoptotic protein expression and downregulating anti-apoptotic protein expression from the BCl-2 protein family. These results demonstrate that higher cellular uptake and reduction of PL-III by biological reductants in PC3 cells resulted in a synergistic effect of lithocholic acid and cisplatin, which can be easily observed due to its unique cytotoxic mechanism. This further underscores the significance of dual-action PtIV complexes in enhancing the efficacy of cancer therapy.

Borneol hinders the proliferation and induces apoptosis through the suppression of reactive oxygen species-mediated JAK1 and STAT-3 signaling in human prostate cancer cells.

The signal transducer and activator of transcription-3 (STAT-3) is a perilous transcription factor that regulates various proliferation and anti-apoptosis factors in prostate cancer cells. Therefore, inhibiting STAT-3 signaling is considered a potential therapeutic approach for treating prostate cancer. This study investigates the effects of borneol (BNL) on the proliferation and apoptosis of human prostate cancer cells by blocking Janus kinase (JAK) and STAT-3 expression. Human prostate cancer (PC-3) cells were treated with varying concentrations of BNL (10, 20, 30 μM) for 24 hours. Subsequent analyses included MTT based cytotoxicity, reactive oxygen species (ROS) production measured by 2,7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, and apoptosis morphological examination was studied by acridine orange/ethidium bromide (AO/EtBR) staining. BNL treatment mediated protein expression of proliferation and apoptosis-related proteins associated with the JAK-STAT-3 pathways was investigated by Western blot. Results of this study indicates that BNL treatment with PC-3 cells induces cytotoxicity, increases ROS production, and causes apoptotic morphological changes in a concentration-dependent manner. BNL significantly reduced the expression of cell proliferation markers such as cyclin-D1, cyclin-D2 and cyclin-E1 (P<0.05) compared to untreated PC-3 control cells. BNL treatment enhanced apoptosis rates by observed overexpression of Bcl-2-associated X protein (Bax), caspase-3 and down regulation B-cell leukemia/lymphoma 2 (Bcl-2) (P<0.05) expression in PC-3 cells. Additionally, BNL reduced interleukin-6, JAK1, and STAT3 phosphorylation ((P<0.05) in PC-3 cells that obstructing the expression of proliferation and anti-apoptotic proteins in PC-3 cells. Thus, BNL may be a therapeutic agent against prostate cancer by blocking the STAT3 signaling axis.

Establishment of BCL-2 Inhibitors-Resistant B-cell Acute Lymphoblastic Leukemia Cell Lines and Study on Their Resistance Mechanisms

RS4;11 cell line was used to establish BCL-2 inhibitor-resistant cell lines of B-cell acute lymphoblastic leukemia (B-ALL) and explore the possible mechanisms of drug resistance. RS4;11 cell line was continuously induced and cultured by low and ascending concentrations of BCL-2 inhibitors navitoclax and venetoclax to construct navitoclax-resistant cell line RS4;11/Nav and venetoclax-resistant cell line RS4;11/Ven. The cell viability was detected by MTT assay, and the cell apoptosis was detected by flow cytometry. Differentially expressed genes (DEGs) between RS4;11 drug-resistant cell lines and parental cell line were detected by transcriptome sequencing technology (RNA-seq), and mRNA expression levels of DEGs between drug-resistant cell lines and parental cell line were detected by real-time PCR (RT-PCR). Western blot was used to detect the expression levels of BCL-2 family anti-apoptotic proteins in drug-resistant cell lines and parental cell line. The drug-resistant cell lines RS4;11/Nav and RS4;11/Ven were successfully established. The resistance index (RI) of RS4;11/Nav to navitoclax and RS4;11/Ven to venetoclax was 328.655±47.377 and 2 894.027±300.311, respectively. The results of cell apoptosis detection showed that compared with the drug-resistant cell lines, RS4;11 parental cell line were significantly inhibited by BCL-2 inhibitors, while the apoptosis rate of drug-resistant cell lines was not affected by the drugs. Western blot assay showed that the expression of anti-apoptotic proteins of BCL-2 family did not increase significantly in drug-resistant cell lines. RNA-seq, RT-PCR and Western blot assays showed that the expression of EP300 in drug-resistant cell lines was significantly higher than that in parental cell line (P <0.05). Drug-resistant B-ALL cell lines could be successfully established by exposing RS4;11 cell line to the ascending concentration of BCL-2 inhibitors, and the drug resistance mechanism may be related to the overexpression of EP300.

Unlocking Apoptotic Pathways: Overcoming Tumor Resistance in CAR-T-Cell Therapy.

Chimeric antigen receptor (CAR)-T-cell therapy has transformed cancer treatment, leading to remarkable clinical outcomes. However, resistance continues to be a major obstacle, significantly limiting its efficacy in numerous patients. This review critically examines the challenges associated with CAR-T-cell therapy, with a particular focus on the role of apoptotic pathways in overcoming resistance. We explore various strategies to sensitize tumor cells to CAR-T-cell-mediated apoptosis, including the use of combination therapies with BH3 mimetics, Mcl-1 inhibitors, IAP inhibitors, and HDAC inhibitors. These agents inhibit anti-apoptotic proteins and activate intrinsic mitochondrial pathways, enhancing the susceptibility of tumor cells to apoptosis. Moreover, targeting the extrinsic pathway can increase the expression of death receptors on tumor cells, further promoting their apoptosis. The review also discusses the development of novel CAR constructs that enhance anti-apoptotic protein expression, such as Bcl-2, which may counteract CAR-T cell exhaustion and improve antitumor efficacy. We assess the impact of the tumor microenvironment (TME) on CAR-T cell function and propose dual-targeting CAR-T cells to simultaneously address both myeloid-derived suppressor cells (MDSCs) and tumor cells. Furthermore, we explore the potential of combining agents like PPAR inhibitors to activate the cGAS-STING pathway, thereby improving CAR-T cell infiltration into the tumor. This review highlights that enhancing tumor cell sensitivity to apoptosis and increasing CAR-T cell cytotoxicity through apoptotic pathways could significantly improve therapeutic outcomes. Targeting apoptotic proteins, particularly those involved in the intrinsic mitochondrial pathway, constitutes a novel approach to overcoming resistance. The insights presented herein lay a robust foundation for future research and clinical applications aimed at optimizing CAR-T cell therapies.