Survival of microsatellite-stable endometrioid endometrial cancer patients after minimally invasive surgery: An analysis of the Cancer Genome Atlas data.

OBJECTIVE: This study aims to investigate the survival impact of minimally invasive surgery on endometrial cancer (EC) patients with different histology and microsatellite status. METHODS: This is a retrospective study based on the Cancer Genome Atlas (TCGA) data. 519 eligible EC patients were divided into four subgroups according to histology and microsatellite status. Kaplan-Meier survival analyses were conducted in all patients and four subgroups to compare the survival outcome after two surgeries (open vs. minimally invasive). Propensity score matching and propensity score covariate adjustment models were used to control confounders. To establish survival prediction models for EC patients, multivariate stepwise Cox regressions were conducted. RESULTS: Among the eligible patients, 318 (61.3%) received open surgery and 201 (38.7%) received minimally invasive surgery. Overall survival was similar between the two groups (p = 0.33), but the latter showed significantly shorter recurrence-free survival (RFS) (p = 0.005). Subgroup analyses revealed the survival influence of surgical approach was only significant in microsatellite-stable (MSS) endometrioid EC patients. These results were verified by Kaplan-Meier survival analyses after propensity score matching and propensity score covariate adjustment models. Finally, the survival influence of multiple clinicopathological factors was analyzed. After stepwise Cox regressions, minimally invasive surgery was found to be independent risk factor for shorter RFS of all patients (hazard ratio [HR] = 2.038, 95% confidence interval [CI] 1.111-3.741, p = 0.02) and MSS patients (HR = 2.449, 95% CI 1.064-5.639, p = 0.04). CONCLUSIONS: Minimally invasive surgery is associated with more rapid recurrence in MSS endometrioid EC patients, thus indicating the necessity of microsatellite testing for guiding EC surgery.

AK-968 efficiently inhibits proliferation and induces cell apoptosis in human multiple myeloma cells.

AK-968 is a small molecular weight compound. In this study, we evaluated its anti-tumor effects on human multiple myeloma (MM) cells. Our results demonstrated that AK-968 markedly inhibited proliferation of MM cell lines in a dose-and time-dependent manner. Additionally, our flow cytometry data showed that MM cell apoptosis was significantly induced by AK-968. As expected, caspase-3 cleavage and alternation of mitochondrial membrane potential (MMP) in the MM cells treated by AK-968 was detected or quantified by Western Blot or Flow cytometry technique, respectively. In conclusion, our results suggested that AK-968 may act as a potential drug to treat human multiple myeloma.

Validation of a one-step genomics-based molecular classifier for endometrial carcinoma in a large Chinese population.

OBJECTIVE: The aim of this study is to delineate the molecular classification features within Chinese endometrial cancer (EC) patients and to evaluate the concurrence between two widely employed methods for diagnosing EC molecular subtypes. METHODS: This retrospective observational cohort study encompassed 479 cases of EC for analysis. Utilizing next-generation sequencing (NGS) panels targeting POLE, TP53, and microsatellite instability (MSI) status, four subtypes [POLE ultramutated (POLE mut), MMR-deficient (MMRd), p53 abnormal (p53abn), and no specific molecular profile (NSMP)] were classified. Immunohistochemistry (IHC) was employed to ascertain the expression of p53 and MMR proteins. RESULTS: Among the 479 patients, the distribution of EC subtypes was as follows: 28 (5.85%) POLE mut, 67 (13.99%) MMRd, 60 (12.53%) p53abn, and 324 (67.64%) NSMP. When compared to published findings on EC subtypes in the Caucasian population, our real-world data on Chinese ECs revealed a notably higher proportion of NSMP/CNL (copy number low). The evaluation of MSI/MMR status through NGS-based and IHC-based methods displayed substantial concordance (Kappa = 0.91). Slight discordance between the two techniques in identifying p53 abnormalities (Kappa = 0.83) might stem from TP53 truncating mutations, cytoplasmic p53 expression, null TP53 mutants, and well-documented challenges in interpreting p53 IHC. CONCLUSIONS: Chinese ECs exhibit distinctive molecular attributes. For accurate molecular subtyping of Chinese ECs, additional molecular markers that align with the Chinese population's characteristics should be incorporated into existing classifiers. The study's outcomes underscore a strong agreement between NGS and IHC in TP53/p53 detection and MSI assessment.

Iron overload promotes the progression of MLL-AF9 induced acute myeloid leukemia by upregulation of FOS.

Systemic iron overload is a common clinical challenge leading to significantly serious complications in patients with acute myeloid leukemia (AML), which affects both the quality of life and the overall survival of patients. Symptoms can be relieved after iron chelation therapy in clinical practice. However, the roles and mechanisms of iron overload on the initiation and progression of leukemia remain elusive. Here we studied the correlation between iron overload and AML clinical outcome, and further explored the role and pathophysiologic mechanism of iron overload in AML by using two mouse models: an iron overload MLL-AF9-induced AML mouse model and a nude xenograft mouse model. Patients with AML had an increased ferritin level, particularly in the myelomonocytic (M4) or monocytic (M5) subtypes. High level of iron expression correlated with a worsened prognosis in AML patients and a shortened survival time in AML mice. Furthermore, iron overload increased the tumor load in the bone marrow (BM) and extramedullary tissues by promoting the proliferation of leukemia cells through the upregulation of FOS. Collectively, our findings provide new insights into the roles of iron overload in AML. Additionally, this study may provide a potential therapeutic target to improve the outcome of AML patients and a rationale for the prospective evaluation of iron chelation therapy in AML.

Molecular pathways and cellular subsets associated with adverse clinical outcomes in overlapping immune-related myocarditis and myositis.

Immune checkpoint therapies (ICTs) can induce life-threatening immune-related adverse events, including myocarditis and myositis, which are rare but often concurrent. The molecular pathways and immune subsets underlying these toxicities remain poorly understood. To address this need, we obtained heart and skeletal muscle biopsies for single-cell RNA sequencing in living patients with cancers treated with ICTs admitted to the hospital with myocarditis and/or myositis (overlapping myocarditis plus myositis, n=10; myocarditis-only, n=1) compared to ICT-exposed patients ruled out for toxicity utilized as controls (n=9) within 96 hours of clinical presentation. Analyses of 58,523 cells revealed CD8+ T cells with a cytotoxic phenotype expressing activation/exhaustion markers in both myocarditis and myositis. Furthermore, the analyses identified a population of myeloid cells expressing tissue-resident signatures and FcγRIIIa (CD16a), which is known to bind IgG and regulate complement activation. Immunohistochemistry of affected cardiac and skeletal muscle tissues revealed protein expression of pan-IgG and complement product C4d that were associated with the presence of high-titer serum autoantibodies against muscle antigens in a subset of patients. We further identified a population of inflammatory IL-1B+TNF+ myeloid cells specifically enriched in myocarditis and associated with greater toxicity severity and poorer clinical outcomes. These results are the first to recognize these myeloid subsets in human immune-related myocarditis and myositis tissues and nominate new targets for investigation into rational treatments to overcome these high-mortality toxicities.

Sox13 and M2-like leukemia-associated macrophages contribute to endogenous IL-34 caused accelerated progression of acute myeloid leukemia.

Interleukin 34 (IL-34) mainly plays physiologic and pathologic roles through the sophisticated multi-ligand signaling system, macrophage colony-stimulating factor (M-CSF, CSF-1)/IL-34-CSF-1R axis, which exhibits functional redundancy, tissue-restriction and diversity. This axis is vital for the survival, differentiation and function of monocytic lineage cells and plays pathologic roles in a broad range of diseases. However, the role of IL-34 in leukemia has not been established. Here MLL-AF9 induced mouse acute myeloid leukemia (AML) model overexpressing IL-34 (MA9-IL-34) was used to explore its role in AML. MA9-IL-34 mice exhibited accelerated disease progression and short survival time with significant subcutaneous infiltration of AML cells. MA9-IL-34 cells showed increased proliferation. In vitro colony forming assays and limiting dilution transplantation experiments demonstrated that MA9-IL-34 cells had elevated leukemia stem cell (LSC) levels. Gene expression microarray analysis revealed a panel of differential expressed genes including Sex-determining region Y (SRY)-box 13 (Sox13). Furthermore, a positive correlation between the expressions of IL-34 and Sox13 was detected human datasets. Knockdown of Sox13 rescued the enhanced proliferation, high LSC level and subcutaneous infiltration in MA9-IL-34 cells. Moreover, more leukemia-associated macrophages (LAMs) were detected in MA9-IL-34 microenvironment. Additionally, those LAMs showed M2-like phenotype since they expressed high level of M2-associated genes and had attenuated phagocytic potential, suggesting that LAMs should also contribute to IL-34 caused adverse phenotypes. Therefore, our findings uncover the intrinsic and microenvironmental mechanisms of IL-34 in AML and broadens the knowledge of M-CSF/IL-34-CSF-1R axis in malignancies.

Enantioselective Oral Absorption of Molecular Chiral Mesoporous Silica Nanoparticles.

Inspired by the unique pharmacological effects of chiral drugs in the asymmetrical body environments, it is assumed that the chirality of nanocarriers is also a key factor to determine their oral adsorption efficiency, apart from their size, shape, etc. Herein, l/d-tartaric acid modified mesoporous silica nanoparticles (l/d-CMSNs) are fabricated via a one-pot cocondensation method, and focused on whether the oral adsorption of nanocarriers will be benefited from their chirality. It is found that l-CMSN performed better in the sequential oral absorption processes, including mucus permeation, mucosa bio-adhesion, cellular uptake, intestinal transport and gastrointestinal tract (GIT) retention, than those of the d-chiral (d-CMSN), racemic (dl-CMSN), and achiral (MSN) counterparts. The multiple chiral recognition mechanisms are experimentally and theoretically demonstrated following simple differential adsorption on biointerfaces, wherein electrostatic interaction is the dominant energy. During the oral delivery task, l-CMSN, which is proven to be stable, nonirritative, biocompatible, and biodegradable, is efficiently absorbed into the blood (1.72-2.05-fold higher than other nanocarriers), and helps the loaded doxorubicin (DOX) to achieve better intestinal transport (2.32-27.03-times higher than other samples), satisfactory bioavailability (449.73%) and stronger antitumor effect (up to 95.43%). These findings validated the dominant role of chirality in determining the biological fate of nanocarriers.

A detailed analysis of lymph node recurrence in endometrial carcinoma.

Background: The lymph node (LN) is one of the main sites of recurrence in patients with endometrial carcinoma (EC). Literature specifically analyzing LN recurrence (LNR) in EC remains limited in number. Methods: Patients with EC undergoing surgery between 2006 and 2021 in Peking University People's Hospital was included, clinicopathological data of whom were collected and analyzed retrospectively by R 4.0.3. Results: A total of 792 patients were included, with 73 patients having recurrence, among whom 21 patients had LNR. Median recurrence-free survival (RFS) in patients with LNR was 16 [4-39] months. LNR was extensive, with pelvic LNs most commonly involved (9/21). There are various patterns of LNR, with 33.3% (7/21) LN-only recurrence. Multivariable analysis suggested advanced stage, larger tumor diameter and poor histology were independent risk factors for LNR. Patients with LN metastasis (LNM) diagnosed at initial treatment accounted for 47.6% (10/21) of cases with LNR, 60.0% (6/10) of whom had recurrent LNs beyond the region of LNM, 90.0% (9/10) of whom had recurrence nodes overlapping with the range of lymphadenectomy. Uni- and multi-variable analysis suggested lymphadenectomy was not a protective factor for LNR, with both the range and number of LNs harvested considered. Conclusions: LNR is common in patients with EC, with an extensive range and various patterns of recurrence. The International Federation of Gynecology and Obstetrics (FIGO) stage, tumor diameter and histology were independent risk factors for LNR, but lymphadenectomy seemed not a protective factor for LNR.

Bone Metastases of Endometrial Carcinoma Treated by Surgery: A Report on 13 Patients and a Review of the Medical Literature.

BACKGROUND: The aim of this study was to describe the clinicopathological features of endometrial cancer (EC) patients with bone metastases treated with surgery and to systematically review the literature. METHODS: We performed a retrospective study to include patients with bone metastases of EC at Peking University People's Hospital from 2000 to 2019. Clinicopathological features and survival outcomes were collected. RESULTS: Among the 1662 patients with EC, 14 (0.84%) were identified with bone metastases, and all were treated surgically. Thirteen cases were analyzed. Four had bone metastases when diagnosed, and the remaining nine cases had bone metastases when first relapsed, with a median time to recurrence of 13 months (range, 5-144). The median age of the 13 patients was 58 years old (range, 45-76). Twelve were endometrioid carcinoma. The majority of sites of bone metastases were the pelvis, followed by the spine. The median overall survival (OS) was 57 months. We further combined the 13 patients with another 24 cases identified from literature research. There was no significant difference in clinicopathological characteristics between the patients with bone metastases when diagnosed and when they first relapsed. The median OS was numerically longer for patients with bone metastases when diagnosed than when they first relapsed (57 vs. 36 months, p = 0.084). CONCLUSIONS: Patients with bone metastases of EC might benefit from comprehensive treatment based on surgery, as symptoms can be palliated and survival can probably be extended.

Loss of IRF7 accelerates acute myeloid leukemia progression and induces VCAM1-VLA-4 mediated intracerebral invasion.

Interferon regulatory factor 7 (IRF7) is widely studied in inflammatory models. Its effects on malignant progression have been documented mainly from the perspective of the microenvironment. However, its role in leukemia has not been established. Here we used MLL-AF9-induced acute myeloid leukemia (AML) mouse models with IRF7 knockout or overexpression and xenograft mouse models to explore the intrinsic effects of IRF7 in AML. AML-IRF7-/- mice exhibited accelerated disease progression with intracerebral invasion of AML cells. AML-IRF7-/- cells showed increased proliferation and elevated leukemia stem cell (LSC) levels. Overexpression of IRF7 in AML cells decreased cell proliferation and LSC levels. Furthermore, overexpression of transforming growth-interacting factor 1 (TGIF1) rescued the enhanced proliferation and high LSC levels caused by IRF7 deficiency. Moreover, upregulation of vascular cell adhesion molecule 1 (VCAM1), which correlated with high LSC levels, was detected in AML-IRF7-/- cells. In addition, blocking VCAM1-very late antigen 4 (VLA-4) axis delayed disease progression and attenuated intracerebral invasion of AML cells. Therefore, our findings uncover the intrinsic effects of IRF7 in AML and provide a potential strategy to control central nervous system myeloid leukemia.

Tumor immune microenvironment in endometrial cancer of different molecular subtypes: evidence from a retrospective observational study.

Objective: Tumor immune microenvironmental features may predict survival and guide treatment. This study aimed to comprehensively decipher the immunological features of different molecular subtypes of endometrial cancer. Methods: In this retrospective study, 26 patients with primary endometrial cancer and four with recurrent disease treated in our center from December 2018 to November 2021 were included. Next-generation sequencing was performed on tumor samples. Patients were classified into four subtypes, including POLE mutant, microsatellite instability high (MSI-H), no specific molecular profile (NSMP) and TP53 mutant subtypes. Tumor-infiltrating immune cells were quantified using multiplex immunofluorescence assays. Results: Of the 26 primary endometrial cancer cases, three were POLE mutant, six were MSI-H, eight were NSMP and nine were TP53 mutant. Of the four recurrent cases, two belonged to the NSMP subtype and two belonged to the TP53 mutant subtype. The tumor mutation burden (TMB) levels of POLE mutant and MSI-H cases were significantly higher than that of the other two subtypes (p< 0.001). We combined POLE mutant and MSI-H subtypes into the TMB high (TMB-H) subtype. The TMB-H subtype showed a high degree of infiltration of CD8+ T cells. In the NSMP subtype, the overall degree of intra-tumoral infiltrating immune cells was low. In the TP53 mutant subtype, the densities of both PD-L1+ macrophages (p = 0.047) and PD-1+ T cells (p = 0.034) in tumor parenchyma were the highest among the four subtypes. Conclusion: Endometrial cancer of TMB-H, NSMP and TP53 mutant subtypes displayed phenotypes of normal immune response, absence of immune infiltration, and suppressed immune response, respectively. These features may provide mechanistic explanations for the differences in patients' prognosis and efficacy of immune checkpoint blockade therapies among different endometrial cancer subtypes.

Microstructure and Wear Resistance of Multi-Layer Ni-Based Alloy Cladding Coating on 316L SS under Different Laser Power.

We prepared three kinds of Ni based alloy cladding coatings on 316L stainless steel at different power levels. The microstructure of the cladding layer was observed and analyzed by XRD, metallographic microscope, and SEM. The hardness of the cladding layer was measured, and the wear resistance of it was tested by a friction instrument. The results show that the effect of laser cladding is good, and it has good metallurgical bonding with the substrate. Different microstructures such as dendritic and equiaxed grains can be observed in the cladding layer. With the increase in laser power, more equiaxed and columnar dendrites can be observed. The phase composition of the cladding layer is mainly composed of γ-Ni solid solution and some intermetallic compounds such as Ni3B, Cr5B3, and Ni17Si3. The results of EDS show that there are some differences in the distribution of C and Si between dendrites. The hardness of the cladding layer is about 600 HV0.2, which is about three times of the substrate (~200 HV0.2). Through the analysis of the wear morphology, the substrate wear is serious, there are serious shedding, mainly adhesive wear, and abrasive wear. However, the wear of the cladding layer is slight, which is abrasive wear, and there are some grooves on the surface.

Sentinel Lymph Node Biopsy Is Feasible in Cervical Cancer Laparoscopic Surgery: A Single-Center Retrospective Cohort Study.

METHODS: A total of 100 cervical cancer patients undergoing laparoscopic surgery with SLN biopsy were included. Indocyanine green, carbon nanoparticles (CNPs), and a combination of both were used during surgeries. Detection rates, sensitivity, negative predictive value (NPV) of SLN biopsy, and related factors were analyzed. RESULTS: The overall and bilateral SLN detection rates were 92% (92/100) and 74% (74/100), respectively. Combined tracers had higher bilateral SLN detection rates than CNPs alone (p=0.005). Menopause and lymph node metastasis were associated with lower overall and bilateral SLN detection rates (p < 0.05). SLN biopsy sensitivity and NPV for lymph node metastasis in patients with at least one detected SLN were 81.8% (9/11) and 97.3% (72/74), respectively. Among those with bilateral detected SLNs, higher sensitivity and NPV of 87.5% (7/8) and 98.3% (57/58) were observed, respectively. SLN algorithm can ensure that all patients with lymph node metastasis are detected by SLN biopsy. CONCLUSION: SLN biopsy appears to be safe and effective for specific cervical cancer patients with high detection rates and NPV in laparoscopic surgery, especially for those with detected bilateral SLNs and undergoing the SLN algorithm. Selecting suitable patients for SLN mapping has prospects for clinical application.

Tumor Molecular Features Predict Endometrial Cancer Patients' Survival After Open or Minimally Invasive Surgeries.

BACKGROUND: The Cancer Genome Atlas (TCGA) project shed light on the vital role of tumor molecular features in predicting endometrial cancer patients' prognosis. This study aims to investigate the survival impact of surgical approaches on patients with different genetic alterations. METHODS: 473 endometrial cancer patients from TCGA database were selected. To analyze the prognostic impact of surgical approach, survival analyses were conducted in patients with different molecular features. Finally, a simplified molecular stratification model was established to select patients suitable for open or minimally invasive surgery (MIS). RESULTS: In our cohort, 291 patients received open surgery and 182 received MIS. Molecular features influenced patients' survival after different surgical approaches. Based on survival analyses, three molecular subtypes were generated, with subtype 1 harboring POLE mutation (POLEmt ), microsatellite-instability high (MSI-H), homologous recombination repair (HRR) pathway mutation or MUC16 mutation (MUC16mt ); subtype 3 carrying TP53 mutation; and subtype 2 without specific molecular feature. The survival influence of molecular subtypes depended on surgical approaches. In the open surgery cohort, three subtypes showed similar survival outcome, while in the MIS cohort, prognosis varied significantly among three subtypes, with subtype 1 the best and subtype 3 the worst. In stepwise Cox regression, molecular subtype was an independent predictor of recurrence-free survival in patients receiving MIS (p < 0.001). CONCLUSION: The molecular features of endometrial cancer are associated with patients' prognosis after different surgical approaches. MIS should be recommended in patients with POLEmt , MSI-H, HRR pathway mutation or MUC16mt , while for patients with TP53 mutation, open surgery is better concerning oncological safety.

Identification of an immune-related risk signature and nomogram predicting the overall survival in patients with endometrial cancer.

OBJECTIVE: Aimed to construct an immune-related risk signature and nomogram predicting endometrial cancer (EC) prognosis. METHODS: An immune-related risk signature in EC was constructed using the least absolute shrinkage and selection operator regression analysis based on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. A nomogram integrating the immune-related genes and the clinicopathological characteristics was established and validated using the Kaplan-Meier survival curve and receiver operating characteristic (ROC) curve to predict the overall survival (OS) of EC patients. The Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) R tool was used to explore the immune and stromal scores. RESULTS: CCL17, CTLA4, GPI, HDGF, HFE2, ICOS, IFNG, IL21R, KAL1, NR3C1, S100A2, and S100A9 were used in developing an immune-related risk signature evaluation model. The Kaplan-Meier curve indicated that patients in the low-risk group had better OS (p<0.001). The area under the ROC curve (AUC) values of this model were 0.737, 0.764, and 0.782 for the 3-, 5-, and 7-year OS, respectively. A nomogram integrating the immune-related risk model and clinical features could accurately predict the OS (AUC=0.772, 0.786, and 0.817 at 3-, 5-, and 7-year OS, respectively). The 4 immune cell scores were lower in the high-risk group. Forkhead box P3 (FOXP3) and basic leucine zipper ATF-like transcription factor (BATF) showed a potential significant role in the immune-related risk signature. CONCLUSION: Twelve immune-related genes signature and nomogram for assessing the OS of patients with EC had a good practical value.

DDIT4 mediates the proliferation-promotive effect of IL-34 in human monocytic leukemia cells.

Interleukin 34 (IL-34) is a cytokine that shares the receptor with colony-stimulating factor 1 (CSF-1). IL-34 is involved in a broad range of pathologic processes including cancer. We previously demonstrated that IL-34 promoted the proliferation and colony formation of human acute monocytic leukemia (AMoL) cells. However, the mechanism has not been elucidated. Here, by analyzing the gene profiles of Molm13 and THP1 cells overexpressing IL-34 (Molm13-IL-34 and THP1-IL-34), upregulation of the DNA damage-inducible transcript 4 (DDIT4) was detected in both series. Knockdown of DDIT4 effectively inhibited the proliferation, promoted apoptosis and colony formation in Molm13-IL-34 and THP1-IL-34 cells. Our results suggest that DDIT4 mediates the proliferation-promotive effect of IL-34 whereas does not mediate the promotive effect of IL-34 on colony formation in AMoL cells.

Prognostic significance of lymphovascular space invasion in patients with endometrioid endometrial cancer: a retrospective study from a single center.

OBJECTIVE: This study aims to analyze factors associated with lymphovascular space invasion (LVSI) and evaluate the prognostic significance of LVSI in Chinese endometrioid endometrial cancer (EEC) patients. METHODS: Five-hundred eighty-four EEC patients undergoing surgery in our center from 2006 to 2016 were selected for analysis. Univariate analysis and multivariate logistic regression were used to examine relevant factors of LVSI. To evaluate the prognostic role of LVSI, survival analyses were conducted. In survival analyses, both multivariate Cox regression and propensity score matching were used to control the confounders. RESULTS: The incidence of LVSI was 12.16% (71/584). Diabetes history (p=0.021), lymph node metastasis (p=0.005), deep myometrial invasion (p<0.001) and negative PR expression (p=0.007) were independently associated with LVSI. Both Kaplan-Meier method and univariate Cox regressions showed LVSI negative and positive cases had similar tumor-specific survival (TSS) and disease-free survival (DFS). After adjusting for the influence of adjuvant therapy and other clinicopathological factors with multivariate Cox regressions, LVSI still could not bring additional survival risk to the patients (p=0.280 and p=0.650 for TSS and DFS, respectively). This result was verified by Kaplan-Meier survival analyses after propensity score matching (p=0.234 and p=0.765 for TSS and DFS, respectively). CONCLUSION: LVSI does not significantly compromise the survival outcome of Chinese EEC patients.

Novel Small Molecular Compound AE-848 Potently Induces Human Multiple Myeloma Cell Apoptosis by Modulating the NF-κB and PI3K/Akt/mTOR Signaling Pathways.

BACKGROUND: We aimed to investigate the anti-multiple myeloma (MM) activity of the new small molecular compound AE-848 (5-bromo-2-hydroxyisophthalaldehyde bis[(1-methyl-1H-benzimidazol-2-yl)hydrazone]) and its underlying anti-MM mechanism. METHODS: Cell viability and apoptosis were detected and quantified by using MTT and flow cytometry, respectively. JC-1 dye-related techniques were used to assess mitochondrial membrane potential (MMP). Western blotting was applied to detect the expression of NF-κB and PI3K/Akt/mTOR pathway-associated proteins. The in vivo activity of AE-848 against MM was evaluated in a MM mouse model. RESULTS: Application of AE-848 into the in vitro cell culture system significantly reduced the viability and induced apoptosis of the MM cell lines, RPMI-8226 and U266, in a dose- and time-dependent manner, respectively. JC-1 dye and Western blotting analysis revealed that AE-848 induced the cleavage of caspase-8, caspase-3, and poly ADP-ribose polymerase (PARP), resulting in loss of mitochondrial membrane potential (MMP). Both the NF-κB and PI3K/AKT/mTOR signaling pathways were involved in AE-848-induced apoptosis of U266 and RPMI8226 cells. Moreover, AE-848 leads to cell cycle arrest of MM cells. Its anti-MM efficacy was further confirmed in a xenograft model of MM. AE-848 administration significantly inhibited MM tumor progression and prolonged the survival of MM-bearing mice. More importantly, our results demonstrated that AE-848 markedly induced primary MM cell apoptosis. CONCLUSION: Our results for the first time showed that the small compound AE-848 had potent in vitro and in vivo anti-myeloma activity, indicating that AE-848 may have great potential to be developed as a drug for MM treatment.

Disulfiram/copper markedly induced myeloma cell apoptosis through activation of JNK and intrinsic and extrinsic apoptosis pathways.

Disulfiram (DSF) is an FDA approved anti-alcoholism drug in use for more than 60 years. Recently, antitumor activity of the DSF/copper (DSF/Cu) complex has been identified. Its anti-multiple myeloma activity, however, has barely been investigated. In the present study, our results demonstrated that the DSF/Cu complex induced apoptosis of MM cells and MM primary cells. The results indicated that DSF/Cu significantly induced cell cycle arrest at the G2/M phase in MM.1S and RPMI8226 cells. Moreover, JC-1 and Western blot results showed that DSF/Cu disrupted mitochondrial membrane integrity and cleaved caspase-8 in MM cells, respectively, suggesting that it induced activation of extrinsic and intrinsic apoptosis pathways. Interestingly, DSF/Cu induced caspase-3 activation was partly blocked by Z-VAD-FMK (zVAD), a pan-caspase inhibitor, indicating at caspase-dependent and -independent paths involved in DSF/Cu induced myeloma cell apoptosis machinery. Additionally, activation of the c-Jun N-terminal kinase (JNK) signaling pathway was observed in DSF/Cu treated MM cells. More importantly, our results demonstrated that DSF/Cu significantly reduced tumor volumes and prolonged overall survival of MM bearing mice when compared with the controls. Taken together, our novel findings showed that DSF/Cu has potent anti-myeloma activity in vitro and in vivo highlighting valuable clinical potential of DSF/Cu in MM treatment.