Transcription Factor E2F7 Activates PKMYT1 to Partially Suppress Adriamycin Sensitivity in Gastric Cancer Through the MAPK Signaling Pathway

ABSTRACT Background: Adriamycin (ADM) resistance remains an obstacle to gastric cancer chemotherapy treatment. Objective: The objective of this study was to study the role and mechanism of transcription factor E2F7 in sensitivity to ADM chemotherapeutic agents in gastric cancer. Methods: Cell viability and cell sensitivity were assessed by CCK-8 and IC50 values of ADM were calculated. The impact of ADM on cellular proliferative capacity was assessed through colony formation assay. The binding relationship between E2F7 and PKMYT1 was then verified by dual luciferase assay and chromatin immunoprecipitation assay. ERK1/ERK2 and p-ERK1/p-ERK2 protein expression levels were detected by western blot. Results: In both gastric cancer tissue and ADM-resistant cells, a conspicuous upregulation of E2F7 and PKMYT1 was observed. Upregulated PKMYT1 was notably enriched in the MAPK signaling pathway. Enhanced levels of E2F7 were shown to not only drive gastric cancer cell proliferation but also engender a reduction in the sensitivity of these cells to ADM. Furthermore, PKMYT1 emerged as a downstream target of E2F7. Activation of E2F7 culminated in the transcriptional upregulation of PKMYT1, and silencing E2F7 reversed the inhibitory impact of PKMYT1 overexpression on ADM sensitivity in gastric cancer cells. Conclusion: E2F7/PKMYT1 axis might promote the proliferation and partially inhibit ADM sensitivity of gastric cancer cells by activating the MAPK pathway.

Cost-effectiveness analysis of apatinib combined with adriamycin for second-line treatment of platinum-resistant recurrent ovarian cancer / 中国药房

OBJECTIVE To evaluate the cost-effectiveness of apatinib combined with adriamycin in the second-line chemotherapy of platinum-resistant recurrent ovarian cancer （OC） from the perspective of the health system in China. METHODS A three-state partitioned survival model was constructed based on the APPROVE clinical trial and related literature data， with a model simulation time frame of 10 years and a 4-week cycle， and both cost and utility values were discounted using a 5% discount rate. Cost and quality-adjusted life years （QALYs） were used as a model output indicator and the incremental cost-effectiveness ratio （ICER） was calculated to evaluate the cost-effectiveness of apatinib combined with adriamycin versus adriamycin chemotherapy in the second-line treatment of platinum-resistant recurrent OC. One-way sensitivity analysis， probability sensitivity analysis and scenario analysis were used to verify the robustness of the base-case analysis results. RESULTS The results of base-case analysis indicated that compared with chemotherapy alone， ICER of patients receiving apatinib combined with adriamycin was 124 678.25 yuan/QALY， which was less than willingness-to-pay （WTP） threshold set in this study [3 times per capita gross domestic product （GDP） of China in 2022 （257 094 yuan）]. The results of scenario analysis showed that， with the extension of the simulation time limit， the ICER of apatinib combined with adriamycin was gradually reduced， and the decline was gradually reduced， but both were less than WTP threshold. The results of single factor sensitivity analysis showed that the factors that had the greatest impact on ICER were the utility value of progression， body surface area， discount rate，and the cost of best supportive treatment， etc. The results of probability sensitivity analysis showed that under WTP threshold set in this study， the economic probability of apatinib combined with adriamycin was about 99%. CONCLUSIONS From the perspective of China’s health system， using three times the per capita GDP in 2022 as the WTP threshold， the combination of apatinib and adriamycin is more cost-effective than adriamycin alone in second-line chemotherapy for platinum-resistant recurrent OC.

Echinacoside hampers malignant progression of breast cancer MCF-7 cells by modulating AKR1B10/ERK signal transduction / 中国中药杂志

This study analyzes the impact of echinacoside(ECH) in the proliferation, metastasis and adriamycin(ADR) resistance of breast cancer(BC) MCF-7 cells via the modulation of aldo-keto reductase family 1 member 10(AKR1B10)/extracellular signal-regulated kinase(ERK) pathway. The chemical structure of ECH was firstly confirmed. MCF-7 cells were treated with different concentration(0, 10, 20, 40 μg·mL~(-1)) of ECH for 48 h. Western blot was used to analyze expression of AKR1B10/ERK pathway-associated proteins and cell counting kit-8(CCK-8) assay to determine cell viability. MCF-7 cells were collected and classified into control group, ECH group, ECH + Ov-NC group, and ECH + Ov-AKR1B10 group. Then Western blot was employed to analyze the expression of AKR1B10/ERK pathway-associated proteins. CCK-8 and 5-ethynyl-2'-deoxyuridine(EdU) assay were used to examine cell proliferation. Cell migration was appraised with scratch assay, Transwell assay, and Western blot. Eventually, MCF-7 cells were treated with ADR for 48 h to induce ADR resistance. Cell viability was tested by CCK-8 assay and cell apoptosis was estimated based on terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL) assay and Western blot. Based on Protein Data Bank(PDB) and molecular docking, the binding affinity of ECH to AKR1B10 was assessed. Various doses of ECH decreased the expression of AKR1B10/ERK pathway-associated proteins in a dose-dependent manner and declined cell viability compared with the control group. Compared with the control group, 40 μg·mL~(-1) ECH blocked the AKR1B10/ERK pathway in MCF-7 cells and inhibited the proliferation, metastasis and ADR resistance of the cells. Compared with the ECH + Ov-NC group, ECH + Ov-AKR1B10 group showed the recovery of some biological behaviors of MCF-7 cells. ECH also targeted AKR1B10. ECH can inhibit the proliferation, metastasis, and ADR resistance of BC cells by blocking AKR1B10/ERK pathway.

Ephedra Herb extract ameliorates adriamycin-induced nephrotic syndrome in rats via the CAMKK2/AMPK/mTOR signaling pathway / 中国天然药物

This study aimed to investigate the effect and mechanisms of Ephedra Herb (EH) extract on adriamycin-induced nephrotic syndrome (NS), providing an experimental basis for the clinical treatment of NS. Hematoxylin and eosin staining, creatinine, urea nitrogen, and kidn injury molecule-1 were used to evaluate the activities of EH extract on renal function. The levels of inflammatory factors and oxidative stress were detected by kits. The levels of reactive oxygen species, immune cells, and apoptosis were measured by flow cytometry. A network pharmacological approach was used to predict the potential targets and mechanisms of EH extract in the treatment of NS. The protein levels of apoptosis-related proteins and CAMKK2, p-CAMKK2, AMPK, p-AMPK, mTOR and p-mTOR in the kidneys were detected by Western blot. The effective material basis of EH extract was screened by MTT assay. The AMPK pathway inhibitor (compound C, CC) was added to investigate the effect of the potent material basis on adriamycin-induced cell injury. EH extract significantly improved renal injury and relieve inflammation, oxidative stress, and apoptosis in rats. Network pharmacology and Western blot results showed that the effect of EH extract on NS may be associated with the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine significantly ameliorated adriamycin-induced NRK-52e cell injury. Methylephedrine also significantly improved the phosphorylation of AMPK and mTOR, which were blocked by CC. In sum, EH extract may ameliorate renal injury via the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine may be one of the material bases of EH extract.

Over-expression of APMAP alleviates glomerular podocyte injury in adriamycin nephropathy / 基础医学与临床

Objective To investigate the effects of adipocyte plasma membrane-associated protein(APMAP)over-expression on glomerular podocyte injury in adriamycin(ADR)nephropathy.Methods The rat model of adriamy-cin nephropathy was constructed by tail vein injection of adriamycin,the expression of APMAP and NF-κB p65 in renal tissue was measured by immunohistochemistry.A mouse glomerular podocytes MPC-5 cell line with APMAP gene over-expression was constructed,then podocyocytes injury model was induced by 0.5 μmol/L ADR and trea-ted with NF-κB signaling pathway activator CU-T12-9.The proliferation of cells was checked by CCK-8.The activity of lactate dehydrogenase(LDH)was determined by ELISA.The apoptosis of podocytes was determined by flow cytometry.Western blot was used to detect protein expressions of NF-κB p65,p-NF-κB p65 and TNF-α.Results APMAP was expressed in kidney tissue of doxorubicin nephropathy rats at a low level,while NF-κB p65 was significantly high expressed(P＜0.05).Over-expression of APMAP increased proliferation of MPC-5 cells and decreased LDH activity,apoptosis rate,and also down-regulated protein expression of NF-κB p65,P-NF-κB p65 and TNF-α under ADR exposure(P＜0.05).However,combined treatment with CU-T12-9 significantly inhibited the ameliorative effect of APMAP over-expression on the damage of MPC-5 cells exposed to ADR.Conclusions The over-expression of APMAP can inhibit ADR-induced glomerular podocyte injury,and its mechanism might be related to the inhibition of NF-κB signaling pathway.

Cytoprotective Effect of Dietary Squalene Supplementation on Experimentally Induced Cardiomyopathy in Rats

Background: Adriamycin is a broadspectrum, potent, older chemotherapy drug and antineoplastic agent used in the treatment of several cancers such as solid tumours, leukaemias, and lymphomas, playing a major role in cancer chemotherapy. Long-term use of this drug results in congestive heart failure and to overcome this effect dietary squalene intake reduces the adverse effects of adriamycin-mediated cardiotoxicity and cellular oxidative stress. Methods: The current study aims to investigate the cytoprotective effects of dietary squalene supplementation on adriamycin-induced cardiomyopathy in rats in terms of alterations in Troponin T, homocysteine, diagnostic marker enzymes, and cardiac tissue histology. Results: The findings show that a 1.5 percent dose of dietary squalene supplementation for 21 days reduced adriamycin-induced changes in homocysteine, troponin T, diagnostic marker enzymes, and lesions in cardiac tissues. Conclusion: The outcomes of the study specified squalene's cytoprotective action which stabilizes membranes against adriamycin-induced oxidative membrane degradation, which is primarily responsible for heart cell irreversible necrosis.

Effect of MiR-155 Knockout Mediated by Dual sgRNAs on Drug Sensitivity of FLT3-ITD+AML / 中国实验血液学杂志

OBJECTIVE@#Two sgRNAs transfected FLT3-ITD+AML cell line MV411 with different binding sites were introduced into CRISPR/cas9 to obtain MV411 cells with miR-155 gene knockout. To compare the efficiency of miR-155 gene knockout by single and double sgRNA transfection and their effects on cell phenotypes.@*METHODS@#The lentiviral vectors were generated containing either single sgRNA or dual sgRNAs and packaged into lentivirus particles. PCR was conducted to measure gene editing efficiency, and miR-155 expression was evaluated by qPCR. CCK-8 assay was used to evaluate the cell proliferation, and calculate drug sensitivity of cells to adriamycin and quizartinib. Annexin V-APC/7-AAD staining was used to label cell apoptosis induced by adriamycin and quizartinib.@*RESULTS@#In the dual sgRNAs transfected cells, a cleavage band could be observed, meaning the success of gene editing. Compared with the single sgRNA transfected MV411 cells, the expression level of mature miR-155-5p was lower in the dual sgRNA transfected cells. And, dual sgRNA transfected MV411 were more sensitive to adriamycin and quizartinib with lower IC50 and higher apoptosis rate.@*CONCLUSION@#The inhibition rate of miR-155 gene expression transfected by dual sgRNA is higher than that by single sgRNA. Dual sgRNA transfection can inhibit cell proliferation, reverse drug resistance, and induce apoptosis more significantly. Compared with single sgRNA transfection, dual sgRNA transfection is a highly efficient gene editing scheme.

Astragalus polysaccharides affects multidrug resistance gene 1 and P-glycoprotein 170 in adriamycin nephropathy rats via regulating microRNA-16/NF-κB axis / 中南大学学报(医学版)

OBJECTIVES@#Nephrotic syndrome is a common disease of the urinary system. The aim of this study is to explore the effect of astragalus polysaccharides (APS) on multidrug resistance gene 1 (MDR1) and P-glycoprotein 170 (P-gp170) in adriamycin nephropathy rats and the underlying mechanisms.@*METHODS@#A total of 72 male Wistar rats were divided into a control group, a model group, an APS low-dose group, an APS high-dose group, an APS+micro RNA (miR)-16 antagomir group and an APS+miR-16 antagomir control group, with 12 rats in each group. Urine protein (UP) was detected by urine analyzer, and serum cholesterol (CHOL), albumin (ALB), blood urea nitrogen (BUN), and creatinine (SCr) were detected by automatic biochemical analyzer; serum interleukin-6 (IL-6), IL-1β, tumor necrosis factor α (TNF-α) levels were detected by ELISA kit; the morphological changes of kidney tissues were observed by HE staining; the levels of miR-16 and MDR1 mRNA in kidney tissues were detected by real-time RT-PCR; the expression levels of NF-κB p65, p-NF-κB p65, and P-gp170 protein in kidney tissues were detected by Western blotting; and dual luciferase was used to verify the relationship between miR-16 and NF-κB.@*RESULTS@#The renal tissue structure of rats in the control group was normal without inflammatory cell infiltration. The renal glomeruli of rats in the model group were mildly congested, capillary stenosis or occlusion, and inflammatory cell infiltration was obvious. The rats in the low-dose and high-dose APS groups had no obvious glomerular congestion, the proliferation of mesangial cells was significantly reduced, and the inflammatory cells were reduced. Compared with the high-dose APS group and the APS+miR-16 antagomir control group, there were more severe renal tissue structure damages in the APS + miR-16 antagomir group. Compared with the control group, the levels of UP, CHOL, BUN, SCr, IL-6, IL-1β, TNF-α, and MDR1 mRNA, and the protein levels of p-NF-κB p65 and P-gp170 in the model group were significantly increased (all P<0.05); the levels of ALB and miR-16 were significantly decreased (both P<0.05). Compared with the model group, the levels of UP, CHOL, BUN, SCr, IL-6, IL-1β, TNF-α, and MDR1 mRNA, and the protein levels of pNF-κB p65 and P-gp170 in the low-dose and high-dose APS groups were significant decreased (all P<0.05); and the levels of ALB and miR-16 were significantly increased (both P<0.05). Compared with APS+miR-16 antagomir control group, the UP, CHOL, BUN, SCr, IL-6, IL-1β, and TNF-α levels, MDR1 mRNA, and the protein levels of p-NF-κB p65 and P-gp170 were significantly increased (all P<0.05). The levels of ALB and miR-16 were significantly decreased in the APS+miR-16 antagomir group compared with the APS+miR-16 antagomir control group (both P<0.05).@*CONCLUSIONS@#APS can regulate the miR-16/NF-κB signaling pathway, thereby affecting the levels of MDR1 and P-gp170, and reducing the inflammation in the kidney tissues in the adriamycin nephropathy rats.

Effects of vinorelbine combined with adriamycin on breast cancer MCF-7 cells and related mechanisms / 中华内分泌外科杂志

Objective:To investigate the in vitro anti-cancer effect of Vinorelbine (NVB) combined with adriamycin (PLD) on human breast cancer MCF-7 cells and related mechanisms.Methods:The effects of NVB and PLD alone or in combination on the proliferation of breast cancer cells were detected by CCK-8 experiment. Flow cytometry was used to detect cell apoptosis and changes in reactive oxygen species (ROS) levels. Western blot experiment was carried out to detect protein expression.Results:The results of CCK-8 showed that compared with the blank control group, the inhibition rates of the vinorelbine treatment group, the adriamycin treatment group and the combined treatment group were 27.6%, 31.2% and 65.4%, compared with the NVB group and PLD group, the difference between the combined treatment group was statistically significant ( P=0.005 vs 0.001) . The results of flow cytometry showed that the proportion of apoptotic cells in each group was 3.54%, 16.95%, 15.01% and 32.24%, compared with the NVB group and PLD group, the difference between the combined treatment group was statistically significant ( P=0.006 vs 0.005) . The levels of reactive oxygen species in each group were 1, 1.03, 1.06 and 1.57, compared with the NVB group and PLD group, the difference between the combined treatment group was statistically significant ( P=0.008 vs 0.007) . Western blot results showed that the expression of p-ERK and p-STAT3 decreased after the combination of NVB and PLD, which inhibited the ERK/STAT3 signaling pathway. Conclusions:The combination of NVB and PLD can promote the apoptosis of breast cancer cells and inhibit the proliferation of breast cancer cells with high efficiency and low toxicity. Its mechanism of action may be related to the up-regulation of ROS levels in cells, thereby inhibiting the activation of the ERK/STAT3 pathway.

Serum metabolomic study of Shenfu decoction on adriamycin-induced cardiomyopathy in mice / 药学实践杂志

Objective To study the reversal effect of Shenfu decoction（SFD）on adriamycin-induced cardiomyopathy and explore its mechanism by using serum metabolomic technology. Methods The BALB/c mouse model of cardiomyopathy induced by adriamycin was established. The corresponding intervention was given. The serum lactate dehydrogenase（LDH）and creatine phosphatase isoenzyme MB（CK-MB）were measured. The ejection fraction (EF) and shortening fraction (FS) were measured by echocardiography. Mouse serum was collected for gas chromatography-mass spectrometry (GC-MS) analysis. The data obtained was analyzed by multivariate and univariate statistical analysis to compare the changes of endogenous metabolites in the serum of mice in the normal group, model group and Shenfu decoction treatment group, to find the potential biomarkers of Shenfu decoction to reverse the adriamycin-induced cardiomyopathy. Metabolic pathway analysis was used to explore the targeted metabolic pathway of Shenfu decoction. Results The levels of serum LDH and CK-MB in the model group were increased significantly, and the values of EF and FS decreased significantly, indicating that the model was successfully established. The above indicators were significantly improved after treatment with Shenfu decoction. 13 potential biomarkers of adriamycin-induced cardiomyopathy were identified by metabonomic analysis, and Shenfu decoction had significant reversal effect on 11 metabolites. Metabolic pathway analysis showed that the synthesis of phenylalanine, tyrosine and tryptophan, arachidonic acid metabolism, phenylalanine metabolism, tricarboxylic acid cycle and dicarboxylic acid metabolism were the main targeted metabolic pathways of Shenfu decoction. Conclusion Shenfu decoction can reverse adriamycin-induced cardiomyopathy by regulating the unbalanced synthesis of phenylalanine, tyrosine and tryptophan, as well as the metabolism of arachidonic acid, phenylalanine, dicarboxylic acid and tricarboxylic acid cycle.

The biological activity of prodigiosin on adriamycin-resistant breast cancer cells in vitro / 中国药理学通报

Aim To investigate the effect of prodigiosin on the proliferation, migration, cell cycle and apopto- sis of adriamycin-resistant breast cancer cell line MCF- 7/ADR.Methods CCK-8, colony formation assay, scratch test and Transwell were used to detect the pro¬liferation and migration of MCF-7/ADR after treatment with different concentrations of prodigiosin.How cy¬tometry and DAP1 staining were used to detect the cell cycle and apoptosis of MCF-7/ADR cell line after treatment with different concentrations of prodigiosin.Western blot was used to detect the effect of prodigiosin on the expression of caspase-3, Bax and Bcl-2 in MCF- 7/ADR cells.Results Prodigiosin inhibited the pro¬liferation and migration of MCF-7/ADR cell line in a concentration- and time-dependent manner ( P <0.05 ).In addition, prodigiosin enhanced the in¬hibitor}' effect of adriamycin on MCF-7/ADR cell line.Prodigiosin arrested MCF-7/ADR cell line cycle in the S phase and induced cell apoptosis in a time- and dose- dependent manner.The percentage of S phase cells treated with 2 mg • L 1 prodigiosin for 48 hours in¬creased to 35.3% , and the apoptotic rate was as high as 64.83% , which were statistically significant com¬pared with the control group ( P < 0.05 ).Prodigiosin could up-regulate the expression of apoptosis protein caspase-3 and Bax and inhibit the expression of Bcl-2 protein in MCF-7/ADK cell.Conclusions Prodigio¬sin can effectively inhibit the proliferation, migration and promote cell apoptosis and regulate cell cycle in adriamvcin-resistant breast cancer cell line MCF-7/ ADR.Prodigiosin can enhance the sensitivity of MCF- 7/ADR cell line to adriamycin.

Protective Effect of Wenyang Huoxue Huatan Prescription on Cardiotoxicity Induced by Anthracyclines / 中国实验方剂学杂志

Objective:To study the protective effect of the Wenyang Huoxue Huatan prescription （WYHXHT） on cardiotoxicity induced by adriamycin. Method:SD rats were randomly divided into the following six groups： a normal control group， an adriamycin model group， a low-dose （4.86 g·kg^-1） WYHXHT group， a middle-dose （9.72 g·kg^-1） WYHXHT group， a high-dose （19.44 g·kg^-1） WYHXHT group， and a dexrazoxane group. Except for the normal control group， the rats in other groups received intraperitoneal injection of 2.5 mg·kg^-1 adriamycin， once a week for six weeks， with a cumulative dose of 15 mg·kg^-1. The normal control group， the adriamycin model group， and the dexrazoxane group received 10 mL·kg^-1 normal saline daily by gavage. In the dexrazoxane group， the rats were subjected to intraperitoneal injection of 25 mg·kg^-1 dexrazoxane 30 min before doxorubicin administration， once a week for six weeks. The general condition of rats was observed and their body weight was monitored. A high-resolution micro-ultrasound imaging system was used to detect rat cardiac function. Hematoxylin-eosin （HE） staining was performed to observe the pathological changes of myocardial tissues of rats. Western blot was used to detect the protein expression of microtubule-associated protein 1 light chain 3 （LC3） Ⅱ， the mammalian homolog of yeast Atg6 （Beclin-1）， and p62 protein in rat myocardial tissues. Result:Compared with the normal control group， rats in the adriamycin model group showed dull fur， reduced food intake and activity， loose stool， low energy， and slow response. Besides， it also displayed reduced body weight （<italic>P</italic><0.01）， decreased left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） （<italic>P</italic><0.01）， myocardial cell degeneration， edema， rupture， and dissolution， expansion of myocardial interstitium， uneven staining of myocardial fiber， visible inflammatory cell infiltration， up-regulated expression of Beclin-1 and LC3Ⅱ in rat myocardial tissues （<italic>P</italic><0.01）， and down-regulated p62 expression （<italic>P</italic><0.01）. Compared with the adriamycin model group， the medium- and high-dose WYHXHT groups exhibited increased body weight， LVEF， and LVFS （<italic>P</italic><0.01）， relieved pathological injury of myocardial tissues， down-regulated expression of LC3Ⅱ and Beclin-1 （<italic>P</italic><0.05，<italic>P</italic><0.01）， and up-regulated expression of p62 （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:WYHXHT can effectively prevent and treat adriamycin-induced cardiotoxicity， and its effect may be related to the inhibition of myocardial cell autophagy. The effect is dominant in the high-dose group.

p-Coumaric acid alleviates adriamycin-induced hepatotoxicity in rats

Objective: To evaluate the effect of p-coumaric acid against adriamycin-induced hepatotoxicity in rats. Methods: The rats were divided into 4 groups. The control group received solvent; the p-coumaric acid group was treated with 100 mg/kg of p-coumaric acid orally for five consecutive days; the adriamycin group was administered with a single dose of adriamycin (15 mg/kg, i.p.), and the p-coumaric acid + adriamycin group was given p-coumaric acid five days before adriamycin administration. Twenty-four hours after the last administration, blood samples were collected for biochemical analysis, and liver tissues were removed for histopathological and immunohistochemistrical studies. Moreover, the levels of tissue lipid peroxidation and enzyme activities of glutathione peroxidase, superoxide dismutase, and catalase in liver tissue were measured. Results: Treatment with p-coumaric acid protected the liver from the toxicity of adriamycin by attenuating the increase in alkaline phosphatase, alanine transaminase, aspartate transaminase, total bilirubin, total cholesterol, triglyceride, and low-density lipoprotein cholesterol and lessening the decrease in high-density lipoprotein cholesterol and albumin. p-Coumaric acid also raised the levels of glutathione peroxidase, superoxide dismutase, and catalase, as well as decreased lipid peroxidation in liver tissue and hepatic IL-1β expression. Additionally, histopathological study confirmed the protective effect of p-coumaric acid against liver damage. Conclusions: p-Coumaric acid can alleviate adriamycin-induced hepatotoxicity.

Immunomodulatory effect of adriamycin in mouse 4T1 model of breast cancer / 生物工程学报

To explore the immunomodulatory effect of adriamycin on 4T1 breast cancer. We used a tandem mass tag-based quantitative proteomic method to detect differential proteins in breast cancer tissues, and multiple bioinformatics databases to analyze the differentially expressed proteins in the proteome. Also, we used enzyme-linked immunosorbent assay to detect the effects of adriamycin on helper T cells 1 and 2 in breast cancer tissues, and flow cytometry to detect CD4+ T cells, CD8+ T cells and regulatory T cells. We discovered the immunomodulatory targets of adriamycin in differential proteins. In total 170 differential proteins were significantly up-regulated, whereas 58 were markedly down-regulated. In addition, 73 proteins were involved in immune regulation. Kyoto encyclopedia of genes and genomes enriched important protein pathways related to cytokines and factor receptors, interleukin 17 pathway and cancer transcriptional regulatory pathways. These pathways and important differential proteins related to immunomodulatory functions were ultimately regulated by adriamycin on CD4+ T cells, CD8+ T cells and regulatory T cells, thereby affecting the prognosis of breast cancer. Moreover, adriamycin significantly increased interleukin 2, CD4+ T and CD8+ T (P<0.01) and markedly reduced regulatory T cells (P<0.05). The function of adriamycin against triple-negative breast cancer was closely related to the immunoregulation process of the differential proteins Ighm, Igkc, S100A8, S100A9 and Tmsb4x. Adriamycin could regulate the content of helper T cells 1 cytokines, CD4+ T and CD8+ T lymphocytes in breast cancer and reduce the number of regulatory T cells to produce immunomodulatory effects.

P-Coumaric acid alleviates adriamycin-induced hepatotoxicity in rats

Objective: To evaluate the effect of p-coumaric acid against adriamycin-induced hepatotoxicity in rats. Methods: The rats were divided into 4 groups. The control group received solvent; the p-coumaric acid group was treated with 100 mg/kg of p-coumaric acid orally for five consecutive days; the adriamycin group was administered with a single dose of adriamycin (15 mg/kg, i.p.), and the p-coumaric acid + adriamycin group was given p-coumaric acid five days before adriamycin administration. Twenty-four hours after the last administration, blood samples were collected for biochemical analysis, and liver tissues were removed for histopathological and immunohistochemistrical studies. Moreover, the levels of tissue lipid peroxidation and enzyme activities of glutathione peroxidase, superoxide dismutase, and catalase in liver tissue were measured. Results: Treatment with p-coumaric acid protected the liver from the toxicity of adriamycin by attenuating the increase in alkaline phosphatase, alanine transaminase, aspartate transaminase, total bilirubin, total cholesterol, triglyceride, and low-density lipoprotein cholesterol and lessening the decrease in high-density lipoprotein cholesterol and albumin. p-Coumaric acid also raised the levels of glutathione peroxidase, superoxide dismutase, and catalase, as well as decreased lipid peroxidation in liver tissue and hepatic IL- 1β expression. Additionally, histopathological study confirmed the protective effect of p-coumaric acid against liver damage. Conclusions: P-Coumaric acid can alleviate adriamycin-induced hepatotoxicity.

Study on relationship between autophagy and drug resistance of leukemia K562/ADM cells / 中国药理学通报

Aim To set up leukemic K562/ADM cells with stable tolerance to 15 fimol • L_1 ADM induced in vitro by long-term and continuous stepwise increment of adriamycin (ADM) concentration, to observe the sensitivity to other chemotherapy drugs and the relationship between autophagy and drug resistance.Methods MTT assay was used to detect the sensitivity of cells to chemotherapy drugs.The morphological changes of autophagy were observed by transmission electron microscope and fluorescence microscopy.Cell apoptosis analysis was performed using Annexin-V/PI double staining and flow cytometry ( FCM ).The expressions of autophagy and drug resistance associated proteins were tested by Western blot.Results K562/ADM cells were cross-resistance to the other chemotherapeu-tics besides adriamyciri, such as pirarubicin, daunoru- bicin, 5-flurouracil, vincristine but not arsenic triox- ide.The number of autophagosomes, the fluorescence intensity of monodansylcadaverine (MDC) and the expression of LC3-H ,Beclin-l in K562/ADM cells were significantly higher than those in K562 cells.The inhibition of autophagy by 3-MA significantly increased the sensitivity of K562/ADM cells to ADM, and 3-MA also effectively inhibited the expressions of drug resistance related proteins P-gp, MRP1 and BCRP in K562/ADM cells.Conclusions The K562/ADM cells resistant to adriamycin occur multidrug resistance, and the drug resistanceis closely related to the level of autophagy.

Quercetin reverses adriamycin resistance by activating GSK-3β/β-catenin pathways through GAS5 in breast cancer cells / 中国临床药理学与治疗学

AIM: To investigate the ability of quercetin to reverse acquire adriamycin (ADR) resistance and explored its probably mechanism. METHODS: The CCK-8 assay was used to detect the cytotoxicity of quercetin in MCF-7/ADR cells and the reversal effect of ADR. The colony formation assay and Hoechst 332582 staining were used to detect the cell proliferation, cell apoptosis and the accumulation of rhodamine 123 (Rh123) respectively. The RNA expression levels of GAS5 and ABCB1 were detected by qRT-PCR. The protein expression levels of GSK-3β, β-catenin, c-MYC, cyclin D1, and ABCB1 were detected by Western blot. RESULTS: Quercetin (10, 20, 40 μmol/L) significantly enhanced the sensitivity of MCF-7/ADR to ADR, inhibitd cell proliferation, and increased the intracellular accumulation of Rh123. Treatment with quercetin in MCF-7/ADR cells, the expression levels of GAS5 and GSK-3β were increased, whereas the expression levels of β-catenin, c-myc, cyclin D1 and ABCB1 were decreased. Further research revealed that reduction of GAS5 by RNA interference (si-GAS5) induced inhibitory effects on the expressions of GAS5 and GSK-3β, and enhanced the expressions of β-catenin, c-myc, cyclin D1, and ABCB1. Furthermore, treatment by quercetin combined with si-GAS5 in MCF-7/ADR cells, the expressions of these proteins were effectively reversed in comparison to quercetin combined with siRNA negative control (sncRNA). CONCLUSION: Quercetin increases the expression of GAS5by GSK-3β/β-catenin signaling pathway, which inhibits the expression of ABCB1, ultimately reversing ADR resistance in the MCF-7/ADR cells.

Effect of Puta on in vitro anticancer activity of Shataputi AbhrakBhasma on lung, leukemia and prostate cancer cell lines

Background: Rasashastra needs to be upgraded using the technological advances, with regards to drugprocessing, development and therapeutics. The potential of Rasaaushadhis need to be explored by subjecting them against newer life threatening diseases like cancer where contemporary medicine haslimitations. Abhrak Bhasma, one of the drugs of Rasashastra, has some peculiar attributes. According toclassical Rasashastra texts, Shataputi Abhrak Bhasma is regarded as a Rasayan, whose efficacy is in directproportion to the number of Putas. Thus increasing number of Putas not only has a significant effect onthe physical, analytical aspects but also the therapeutic effect of the Abhrak Bhasma.Objectives: To screen in vitro anticancer activity of Abhrak Bhasma at various stages of Putas (20, 50, 100).To evaluate and thus validate the principle from classical Rasashastra texts, which explains direct relationof number of Putas with therapeutic efficacy.Materials and methods: Shataputi Abhrak Bhasma, at various stages of its preparation was subjected toin vitro anticancer activity on three different cancer cell lines (LungHOP62, LeukemiaU937, ProstateDU145) at Tata Memorial Centre- Advanced Centre for Treatment, Research Education in Cancer, NaviMumbai. SRB assay was followed to evaluate the anti-proliferative activity.Results: It was found that Abhrak Bhasma shows concentration dependent positive in vitro anticanceractivity on all three cell lines with highly significant activity on prostate cancer cell lines. Anticanceractivity of Abhrak Bhasma is in the order 100 Puti > 50 Puti > 20 Puti. Shataputi Abhrak Bhasma hadmaximum activity on prostate cancer cell lines almost equivalent to positive control drug adriamycin.Conclusion: The in vitro anticancer activity of Shataputi Abhrak Bhasma increases with increasing numberof Putas, thus revalidating the direct relation between number of Putas and efficacy of the drug.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V

Effect of New Shengmai Decoction on apoptosis and protein expression of Bax, Bcl-2, and Caspase-3 of cardiomyocyte induced by adriamycin / 中草药

Objective: To observe the reversal and prevention effect of New Shengmai Decoction on the rats’ cardiomyocyte apoptosis induced by adriamycin, and provide the experimental foundation basis for the clinical treatment of myocardial injury induced by adriamycin. Method: The rat models with cardiomyocyte apoptosis were established by adriamycin. Forty male SD rats were divided randomly into four groups, including the normal group, the adriamycin model group, the captopril group and the New Shengmai Decoction group. During the experiment, the mental state, activity, feeding, hair color and other conditions of the rats were observed. After 6 weeks of treatment, the cardiac function and left ventricular hypertrophy of rats were measured and the pathological changes of myocardium were observed by HE staining. And the apoptosis of myocardial cells was observed by TUNEL staining and protein expressions of Caspase-3, Bcl-2, and Bax were detected by immunohistochemistry. Results: Compared with the control group, the cardiac function of the model group was significantly decreased. Compared with the model group, the cardiac function of rats after the different drugs’ treatment could be improved in the different degrees and the effect of the New Shengmai Decoction group was significant. Compared with the control group, the heart body ratio, left ventricular hypertrophy index and myocardial cell apoptosis index in the model group were all significantly increased. Compared with the model group, the captopril group and the New Shengmai Decoction group ameliorated cardiac hypertrophy and myocardial cell apoptosis in rats. Compared with the control group, the expression level of bcl-2 protein in the model group was decreased, while the expression level of Bax and Caspase-3 protein was increased. Compared with the model group, captopril and the New Shengmai Decoction increased the expression level of bcl-2 protein and decreased the expression level of Bax and Caspase-3 protein. Conclusion: The New Shengmai Decoction can improve the cardiac function and lessen cardiomyocyte apoptosis of rats. It can also decrease the expression of protein Caspase-3 in the cardiac muscle of rats or inhibit its activity. In order to restrain cardiomyocyte apoptosis, the New Shengmai Decoction can increase the expression of protein Bcl-2 and decrease the expression of protein Bax through improving the expression of Bcl-2/Bax in the cardiac muscle of rats.

Mechanism of celecoxib reverses adriamycin resistance in NK/T cell lymphoma cells by Notch 1/NF-κB/STAT3 pathway / 中国临床药理学与治疗学

AIM: To observe the mechanism of celecoxib reversal adriamycin resistance in NK/T cell lymphoma cells. METHODS: SNK6 and SNK6/ADR cells were treated with celecoxib of different concentrations (10, 20, 40, 60, 80, 100 μmol/L), the growth inhibition rate of SNK6 and SNK6/ADR were measured by MTT method. The IC