Synergistic Antitumor Effect of Everolimus Combined with Gemcitabine on Diffuse Large B-Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of mTOR inhibitors everolimus (EVE) and gemcitabine (GEM) on the proliferation, apoptosis and cell cycle of diffuse large B-cell lymphoma (DLBCL) cell line U2932, and further explore the molecular mechanisms, so as to provide new ideas and experimental basis for the clinical treatment of DLBCL.@*METHODS@#The effect of EVE and GEM on the proliferation of U2932 cells was detected by CCK-8 assay, the IC50 of the two drugs was calculated, and the combination index (CI=) of the two drugs was calculated by CompuSyn software. The effect of EVE and GEM on apoptosis of U2932 cells was detected by flow cytometry with AnnexinV-FITC/PI staining. Flow cytometry with propidium iodide (PI) staining was used to detect the effect of EVE and GEM on the cell cycle of U2932 cells. Western blot assay was used to detect the effects of EVE and GEM on the channel proteins p-mTOR and p-4EBP1, the anti-apoptotic proteins MCL-1 and Survivin, and the cell cycle protein Cyclin D1.@*RESULTS@#Both EVE and GEM could significantly inhitbit the proliferation of U2932 cells in a time- and dose-dependent manner (r=0.465, 0.848; 0.555, 0.796). According to the calculation of CompuSyn software, EVE combined with GEM inhibited the proliferation of U2932 cells at 24, 48 and 72 h with CI=<1, which had a synergistic effect. After treated U2932 cells with 10 nmol/L EVE, 250 nmol/L GEM alone and in combination for 48 h, both EVE and GEM induced apoptosis, and the difference was statistically significant compared with the control group (P<0.05). The apoptosis rate was significantly enhanced after EVE in combination with GEM compared with single-agent (P<0.05). Both EVE and GEM alone and in combination significantly increased the proportion of cells in G1 phase compared with the control group (P<0.05). The proportion of cells in G1 phase was significantly increased when the two drugs were combined (P<0.05). The expression of p-mTOR and effector protein p-4EBP1 was significantly downregulated in the EVE combined with GEM group, the expression of anti-apoptotic proteins MCL-1, Survivin and cell cycle protein cyclin D1 was downregulated too (P<0.05).@*CONCLUSION@#EVE combined with GEM can synergistically inhibit the proliferation of U2932 cells, and the mechanism may be that they can synergistically induce apoptosis by downregulating the expression of MCL-1 and Survivin proteins and block the cell cycle progression by downregulating the expression of Cyclin D1.

The dual mTORC1/2 inhibitor AZD2014 inhibits acute graft rejection in a rat liver transplantation model / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effect of AZD2014, a dual mTORC1/2 inhibitor, against acute graft rejection in a rat model of allogeneic liver transplantation.@*METHODS@#Liver transplantation from Lewis rat to recipient BN rat (a donor-recipient combination that was prone to induce acute graft rejection) was performed using Kamada's two-cuff technique. The recipient BN rats were randomized into 2 groups for treatment with daily intraperitoneal injection of AZD2014 (5 mg/kg, n=4) or vehicle (2.5 mL/kg, n=4) for 14 consecutive days, starting from the first day after the transplantation. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBIL) levels of the rats were measured 3 days before and at 1, 3, 5, 7, 10, and 14 days after the transplantation, and the survival time of the rats within 14 days were recorded. Immunohistochemical staining was used to examine the expressions of CD3 and Foxp3 in the liver graft, and acute graft rejection was assessed using HE staining based on the Banff schema.@*RESULTS@#Three rats in the control group died within 14 days after the surgery, while no death occurred in the AZD2014 group, demonstrating a significantly longer survival time of the rats in AZD2014 group (χ2=4.213, P=0.04). Serum ALT, AST and TBIL levels in the control group increased progressively after the surgery and were all significantly higher than those in AZD2014 group at the same time point (P < 0.05). Pathological examination revealed significantly worse liver graft rejection in the control group than in AZD2014 group based on assessment of the rejection index (P < 0.01); the rats in the control group showed more serious T lymphocyte infiltration and significantly fewer Treg cells in the liver graft than those in AZD2014 group (P < 0.01).@*CONCLUSIONS@#AZD2014 can effectively inhibit acute graft rejection in rats with allogeneic liver transplantation.

Expert consensus on the clinical application of PI3K/AKT/mTOR inhibitors in the treatment of advanced breast cancer / 中华肿瘤杂志

Phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway (PAM pathway) plays an important role in the development of breast cancer and are closely associated with the resistance to endocrine therapy in advanced breast cancer. Therefore, anti-cancer treatment targeting key molecules in this signaling pathway has become research hot-spot in recent years. Randomized clinical trials have demonstrated that PI3K/AKT/mTOR inhibitors bring significant clinical benefit to patients with advanced breast cancer, especially to those with hormone receptor (HR)-positive, human epidermal growth factor receptor (HER) 2-negative advanced breast cancer. Alpelisib, a PI3K inhibitor, and everolimus, an mTOR inhibitor, have been approved by Food and Drug Administration. Based on their high efficacy and relatively good safety profile, expanded indication of everolimus in breast cancer have been approved by National Medical Products Administration. Alpelisib is expected to be approved in China in the near future. The members of the consensus expert panel reached this consensus to comprehensively define the role of PI3K/AKT/mTOR signaling pathway in breast cancer, efficacy and clinical applications of PI3K/AKT/mTOR inhibitors, management of adverse reactions, and PIK3CA mutation detection, in order to promote the understanding of PI3K/AKT/mTOR inhibitors for Chinese oncologists, improve clinical decision-making, and prolong the survival of target patient population.

Actualización de medicamentos asociados a necrosis avascular de los maxilares: perspectiva y revisión de literatura / Update of medications associated with avascular necrosis of the jaws: a perspective and literature review

La osteonecrosis de los maxilares está definida como la exposición de hueso necrótico en la región maxilofacial al menos por ocho semanas en pacientes que están recibiendo medicamentos antirresortivos para el tratamiento del cáncer primario o metastásico hacia el hueso, osteoporosis o enfermedad de Paget, sin historia previa de radiación. Desde el año 2003, la terminología utilizada estaba en relación con los bifosfonatos, en la actualidad ha sido introducido el término osteonecrosis de los maxilares relacionada por medicamentos (OMAM). La cirugía oral (implantología o cirugía periapical) incrementa el riesgo de OMAM, así como los desbalances concomitantes de la salud oral (inflamación dental y formación de abscesos). Las estrategias conservadoras en el tratamiento varían desde el cuidado local conservador hasta la resección quirúrgica radical del hueso necrótico. En el presente artículo se expone un análisis sistemático retrospectivo de la literatura en páginas como PubMed, ScienceDirect y Springer, Cochrane Library. Con el objetivo de resaltar el aumento de la incidencia de OMAM a nivel mundial con el uso de antirresortivos y otros medicamentos asociados en su patogenia en el Hospital Regional «General Ignacio Zaragoza¼ del ISSSTE, UNAM, en la Ciudad de México (AU)

Osteonecrosis of the jaws is defined as the exposure of necrotic bone in the maxillofacial region for at least 8 weeks in patients receiving antiresorptive medications for the treatment of primary or metastatic cancer towards the bone, osteoporosis, or Paget's disease, without previous history of radiation. Since 2003, the terminology used was related to bisphosphonates, the term medication-related osteonecrosis of the jaws has now been introduced. Oral surgery (implantology or periapical surgery) increases the risk of avascular necrosis, as well as concomitant imbalances in oral health (dental inflammation and abscess formation). Conservative strategies in treatment vary from conservative local care to radical surgical resection of the necrotic bone. In this article, a systematic retrospective analysis of the literature is presented on pages such as PubMed, Science Direct and Springer, Cochrane Library. And in which the objective is to highlight the increase in the incidence of medication related osteonecrosis of the jaws worldwide with the use of antiresorptive, and other associated medications in its pathogenesis at the Hospital Regional «General Ignacio Zaragoza¼ ISSSTE, UNAM in Mexico City (AU)

Research progress on risk factors of de novo malignancy after liver transplantation / 器官移植

De novo malignancy after liver transplantation is an important factor that affecting the long-term survival of recipient. The main risk factors for de novo malignancy include immunosuppression and many factors of recipients, such as age, gender, race, primary disease, preoperative tumor history and precancerous lesion, carcinogenic virus infection, smoking and drinking, etc. Currently, there is no standardized monitoring scheme after liver transplantation, but planned monitoring is required for high-risk recipients, thus to achieve early diagnosis and improve the survival rate. This article summarized the incidence, prognosis and related risk factors of de novo malignancy after liver transplantation, which provided reference for improving long-term survival rate of recipients after liver transplantation.

Synthesis and syntehtic process optimization of mTOR protein inhibitor Torin2 / 国际药学研究杂志

Objective: To synthesize the mammalian target of sirolimus(mTOR)inhibitor Torin2 and optimize the synthetic process. Methods: Starting from p-bromoaniline, the target product was obtained through amino bonding, cyclization, chlorination, bonding with 3-aminotoluene ring, oxidation, cyclization and Suzuki reaction. The synthetic conditions for the key intermediates 3 and 8 as well as the final product Torin2 were optimized by the orthogonal experiment. Results and Conclusion: The structures of the intermediates and the target compound were confirmed by MS and 1H NMR data. The total yield of the target compound synthesis increased from 3% to 18%, and no column chromatography was required for the target compound separation and purification, which thus makes it suitable for industrial production.

Dual mTORC1/2 inhibitor AZD2014 inhibits proliferation of HCCLM3 cells through induction of autophagy / 解放军医学杂志

Objective To investigate the effect of mTORC1/2 inhibitor AZD2014 induced autophagy on cell proliferation in HCCLM3 cell lines in vitro. Methods HCCLM3 cells were cultured in the presence or absence of AZD2014 (100 nmol/L) or rapamycin (100 nmol/L) or 3-methyladenine (10 nmol/L). To detect autophagy vacuoles, HCCLM3 cells were divided into the control group, rapamycin group, AZD2014 group, control+3-MA group, rapamycin+3-MA group, and AZD2014+3-MA group, and auto-fluorescent dye monodansylcadaverine (MDC) was used to monitor autophagy vacuoles. To semi-quantify the expression of autophagy-related proteins, HCCLM3 cells were divided into the control group, rapamycin group, and AZD2014 group, and Western blotting analysis was carried out to detect the expression of autophagy-related proteins, LC3B-II and Beclin-1. To evaluate the effect of autophagy induced by AZD2014 on cell proliferation, HCCLM3 cells were divided into the control group, 3-MA group, AZD2014 group, and AZD2014+3-MA group, Cell Counting Kit-8 was used. Results AZD2014-treated HCC cells showed an increase in the number of MDC-labeled vacuoles as well as in their size. AZD2014 treatment increased the levels of LC3B-II and Beclin-1 (P<0.05). CCK-8 assay revealed that suppression of cell proliferation elicited by AZD2014 in HCCLM3 was partly attenuated by 3-MA (autophagy inhibitor). Conclusion Dual mTORC1/2 inhibitor AZD2014 suppressed cell proliferation in the HCCLM3 cell line with increased autophagy, in vitro. This study provides a potential basis for further investigation of AZD2014 as a clinical molecular targeted agent for HCC treatment.

Apatinib Combined with CCI-779 Inhibits the Proliferation and Migration of Small Cell Lung Cancer NCI-H446 Cells In Vitro / 中国肺癌杂志

BACKGROUND@#Lung cancer is the most common malignancy world-wide. Small cell lung cancer is the deadliest subtype of lung cancer, which features such as rapid growth, early metastasis, and high vascularization. Apatinib is a vascular endothelial growth factor receptor 2 inhibitor independently developed in China, which has a significant inhibition in a variety of solid tumors. The purpose of this study is to investigate the effects of Apatinib alone or Apatinib combined with mammalian target of rapamycin (mTOR) inhibitor, CCI-779, on small cell lung cancer cell line NCI-H446 in vitro.@*METHODS@#The small cell lung cancer cell line NCI-H446 was grew in vitro. The effects of Apatinib alone or Apatinib combined with CCI-779 on proliferation, apoptosis, cell cycle and migration of NCI-H446 small cell lung cancer cells were detected by CCK8; FACS and transwell assays were also carried out; Western blot assays were used to detect vascular endothelial growth factor and cell cycle related protein expression.@*RESULTS@#CCK8 assays showed that high concentration of Apatinib could inhibit the proliferation of NCI-H446 cells. Apoptosis assays showed that high concentration of Apatinib could induce NCI-H446 cell apoptosis. Transwell assays showed that high concentration of Apatinib could inhibit NCI-H446 cell migration. After combined with mTOR inhibitor CCI-779, low concentration of Apatinib could inhibit the proliferation and migration of NCI-H446 small cell lung cancer cells and induce apoptosis.@*CONCLUSIONS@#Apatinib has a concentration-dependent effect on the small cell lung cancer cell line NCI-H446. High concentration of Apatinib can inhibit the proliferation and migration of NCI-H446 small cell lung cancer cells, induce apoptosis. Apatinib combined with the mTOR inhibitor CCI-779 can sensitize the NCI-H446 cells to Apatinib.

Rapamycin inhibiting the progression of acute myeloid leukemia with DNMT3A mutation / 上海交通大学学报（医学版）

Objective • To investigate the effect of mammalian target of rapamycin (mTOR) inhibitor rapamycin on acute myeloid leukemia (AML) with DNA methyltransferase 3A (DNMT3A) R882 mutation in mouse model. Methods • AML cell line OCI-AML3 cells with DNMT3A R882 mutation were cultured with the treatment of rapamycin or DMSO, and then these cells were injected into the tail vein of sublethally irradiated NOD/SCID mice, respectively. The disease progression was monitored by blood routine examination and flow cytometry analysis of CD45+, OCI-AML3 cells, in peripheral blood. Survival time was recorded. Samples from bone marrow, spleen and liver were harvested for flow cytometry analysis and pathological examination. Results • The increasing trend of peripheral leukocytes in the rapamycin treated group was obviously slower than that in the DMSO treated group. The proportion of peripheral blood CD45+ cells in the rapamycin treated group was (4.44±2.58)% (1 week after transplantation) and (34.42±13.64)% (2 weeks after transplantation), which were lower than (16.71±8.96)% and (51.55±5.36)% in the DMSO treated group in the same period, respectively. The median survival time of the rapamycin treated group (27 d) was significantly longer than that of the DMSO group (23 d). The ratios of CD45+ cell infiltration in bone marrow, spleen and liver of the rapamycin treated group were all less than 5%, which were significantly lower than those [(51.32±27.81)% in spleen and (50.03±28.74)% in liver] of the DMSO treated group. Compared with the DMSO treated group, the spleen size of the mice was significantly smaller, and the spleen infiltration and structural damage were significantly alleviated in the rapamycin treated group. Conclusion • Rapamycin shows effective inhibition on the progression of AML with DNMT3A R882 mutation in NOD/SCID mouse model.

Synthesis of ATP-competitive inhibitor Wye-125132 / 军事医学

Objective To synthesize Wye-125132,an inhibitor of mTOR,and to establish a synthetic route for industrial production.Methods Barbituric acid was used as the raw material to synthesize the intermediate 2, 4, 6-trichrolo-pyrimidine-5-carbaldehyde 2 via Vilsmeier-Haack and chlorination reaction, while intermediate 5 was prepared via 3-step reaction from 1,4-dioxaspiro-[4,5]decan-8-one.The condensation of 2 and 5 was followed by substitution reaction to obtain the key intermediate 7.The side chain 9 was prepared by 3-step reaction from bromine aniline.Then the title product 1 was obtained with the reaction of Suzuki cross-coupling of 7 and 9.The structures of intermediate and target compounds were confirmed by MS and 1 H-NMR.Results and Conclusion Compared with the method reported in the literature,this new synthesis method possesses some advantages, such as ready availablity of raw materials, simple operation, mild reaction conditions and easy disposal of products.The total yield is 13.2%,and the purity of the target compound is 99.77%.

Progress of research on mTOR inhibitor in endocrine and HER-2 targeted therapy resistance in breast cancer / 国际肿瘤学杂志

Mammalian target of rapamycin(mTOR)locates at the downstream of phosphatidylinositol 3 kinase(PI3K)-protein kinase B(Akt)cell signal transduction pathway. Studies find that the abnormal activa-tion of this pathway is correlated with the endocrine and drug resistance of anti human epidermal growth factor receptor-2(HER-2)target therapy in breast cancer. The combination with mTOR inhibitors based on the past traditional drugs can block the pathway and reflect a favourable application prospect in preventing the develop-ment of resistance and restoring the initial sensitivity on tumor cells. mTOR inhibitors are expected to be the new hope for the treatment of breast cancer.

Research on mTOR inhibitors used in radio-sensitization / 中国肿瘤临床

Radiotherapy is important for cancer treatment. However, some patients still experience relapse and exhibit radiation resistance. Mammalian target of rapamycin (mTOR) is the main effector molecule in PI3K/AKT signaling. This molecule is found in two structurally and functionally distinct multi-protein complexes known as the mTOR complex 1 and mTOR complex 2. The mTOR signaling pathway controls the growth, proliferation, survival, and apoptosis of cancer cells. This pathway is closely related to tumori-genesis and treatment response, and is used in sensitizing radiotherapy. mTOR inhibitors regulate radio-sensitization through multiple mechanisms, including cell cycle alterations, DNA repair modulation, and tumor hypoxia reduction. Preclinical studies showed that mTOR inhibitors with tolerable toxicity may be used as an effective modality to overcome radio-resistant tumors. Responses to mTOR inhibitors vary depending on the cell lines. Molecular markers can be used to select suitable patients. Further studies are needed to com-pletely understand the use of mTOR inhibitors in radio-sensitization.

Expression of mTOR in breast cancer and the effect of mTOR inhibitor on the cell prolofera-tion and apoptosis of breast cancer cells / 临床与实验病理学杂志

Purpose To investigate the expression of mTOR in breast cancer, and to observe the effect of CCI-779 on proliferation and apoptosis of MDA-MB-231 cell. Methods Immunohistochemical staining was used to detect the expression of mTOR protein in breast cancer tissue and MDA-MB-231 cell. MTT method was used to show effect of CCI-779 on proliferation of MDA-MB-231 cell. Annex-inV-FITC/PI method was used to show effect of CCI-779 on apoptosis of MDA-MB-231 cell. Results 54.9% in 71 cases of breast cancer tissue could express mTOR protein, the expression was significantly higher than in 32 cases of normal tissue (21.9%), mTOR protein was also detected in MDA-MB-231 cell, CCI-779 could inhibit the proliferation of MDA-MB-231 cell, and show a dose-and time-dependent, but CCI-779 could not induce apoptosis of MDA-MB-231 with AnnexinV-FITC/PI assay. Conclusion mTOR is closely related to the formation of breast cancer, CCI-779 has strong activity against MDA-MB-231 cell, it has prospect for treatment of breast cancer in the future.

Advancements in mTOR inhibitors for the management of hormone receptor-positive breast cancer / 中国肿瘤临床

Therapies targeting endocrine receptors and human epidermal growth factor receptor 2 have become important treat-ment modes for patients with hormone receptor breast cancer. Despite the availability of these options, however, development of prima-ry or secondary drug resistance and subsequent disease progression in patients with advanced disease continue to occur. Mammalian tar-get of rapamycin (mTOR), a key regulator of cell growth and proliferation, has been implicated in the induction of cellular processes leading to the uncontrolled growth of cancer cells. Recent studies have suggested that overactivation of the mTOR pathway may be in-volved in the development of endocrine resistance. Interrupting this signaling cascade may alleviate such resistance and help restore drug sensitivity. A number of agents targeting the mTOR pathway have shown potent anti-tumorigenic effects in vitro, and several agents show great potential for treating breast cancer patients. Many clinical studies have shown that combining endocrine therapy with mTOR inhibitors could significantly increase the survival rate of breast cancer patients. In this study, we focus on recent research prog-ress on mTOR and its inhibitors in endocrine therapy resistance in breast cancer.

mTOR and glomerular disease / 国际儿科学杂志

The mammalian target of rapamycin (mTOR)is a evolutionary conserved serine/threonine kinase that catalytics specific substrates and regulates cellular basic physiological events,such as growth,proliferation,autophagy and protein translation.It plays an important role in the regulation of cancer,diabetes and a variety of kidney diseases.Most of glomerular diseases are resulted from the podocytes and filtration barrier damage.Studies have suggested that there is a close relationship between mTOR and podosytes damage,but specific mechanism of regulation is not clear now.This review discusses the regulatory mechanism of mTOR kinase and its role in the glomerular disease to provide new ideas for understanding of glomerular disease pathogenesis.

mTOR signaling pathway and colorectal cancer / 国际肿瘤学杂志

Mammalian target of rapamycin (mTOR) signaling pathway,one of the most important cellular signaling transduction pathways,regulates cell survival,proliferation,transdifferentiation,migration and cell cycle by influencing the activation state of various effector molecules in the downstream.Abnormal regulations of these machanisms are closely related to the tumorigenesis and progression of colorectal cancer (CRC).Currently,mTOR inhibitors have been approved in clinical trials of CRC and have made some progress.

Mammalian target of rapamycin inhibitors for treatment in tuberous sclerosis / 소아과

Tuberous sclerosis complex (TSC) is a genetic multisystem disorder that results from mutations in the TSC1 or TSC2 genes, and is associated with hamartomas in several organs, including subependymal giant cell tumors. The neurological manifestations of TSC are particularly challenging and include infantile spasms, intractable epilepsy, cognitive disabilities, and autism. The TSC1- and TSC2-encoded proteins modulate cell function via the mammalian target of rapamycin (mTOR) signaling cascade, and are key factors in the regulation of cell growth and proliferation. The mTOR pathway provides an intersection for an intricate network of protein cascades that respond to cellular nutrition, energy levels, and growth factor stimulation. In the brain, TSC1 and TSC2 have been implicated in cell body size, dendritic arborization, axonal outgrowth and targeting, neuronal migration, cortical lamination, and spine formation. The mTOR pathway represents a logical candidate for drug targeting, because mTOR regulates multiple cellular functions that may contribute to epileptogenesis, including protein synthesis, cell growth and proliferation, and synaptic plasticity. Antagonism of the mTOR pathway with rapamycin and related compounds may provide new therapeutic options for TSC patients.

Advance in research on the relationship between mTOR signaling pathway and tumors / 西安交通大学学报(医学版)

Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase that is centrally involved in the control of cell growth, proliferation, differentiation and cell cycle regulation. Recent studies have found that the mTOR pathway is complicated and the dysregulation of mTOR pathway is recognized to be associated with lots of tumors. Several inhibitors of mTOR could inhibit oncogene transformation, growth and angiogenesis of tumors. So far, four inhibitors of mTOR have been tested in clinical trials, and some have been accessed to phase Ⅱ or Ⅲ trial. Therefore, in this review, we discuss the regulators that govern mTOR pathway activity, and highlight clinical results obtained with the first generation of mTOR inhibitors to reach the oncology clinics.

Malignancy in Renal Transplant Recipients / 대한이식학회지

With improved management of infectious and cardiovascular complications of renal transplant recipients, prolonged survival with long-term follow up duration, and increasing age of patients, cancer became an increasingly important cause of morbidity and mortality in transplant patients. Literatures indicate three to fivefold incidence of malignancy in solid organ transplant recipients compared with that of general population. Certain types of malignancy, such as skin cancer, Kaposi's sarcoma, lymphoma, cervical, oral and anogenital cancer, hepatocelluar and renal cell carcinoma are particularly reported to be high in incidence. Reduction of immunosuppressive medication is the first step to be considered for the management, especially for virus-associated cancer. The inhibitor of mammalian target of rapamycin(mTORi) suppresses the growth and proliferation of tumors in various animal models, cured Kaposi's sarcoma and seems to reduce the incidence of de novo malignancies of renal transplant recipients, and is therefore a valuable option for the management of cancer of these patients without increment of the risk of graft rejection.