Dihydromyricetin reduces lipid accumulation in LO2 cells via AMPK/mTOR-mediated lipophagy pathway and inhibits HepG2 cell proliferation in vitro / 南方医科大学学报

OBJECTIVE@#To explore the mechanism underlying the hepatoprotective effect of dihydromyricetin (DMY) against lipid accumulation in light of the lipophagy pathway and the inhibitory effect of DMY on HepG2 cell proliferation.@*METHODS@#LO2 cells were cultured in the presence of 10% FBS for 24 h and treated with 100 μg/mL DMY, or exposed to 50% FBS for 24 h followed by treatment with 50, 100, or 200 μg/mL DMY; the cells in recovery group were cultured in 50% FBS for 24 h and then in 10% FBS for another 24 h. Oil red O staining was used to observe the accumulation of lipid droplets in the cells, and the levels of TC, TG, and LDL and activities of AST, ALT and LDH were measured. The expression of LC3 protein was detected using Western blotting. AO staining and transmission electron microscopy were used to determine the numbers of autophagolysosomes and autophagosomes, respectively. The formation of autophagosomes was observed with MDC staining, and the mRNA expression levels of LC3, ATG7, AMPK, mTOR, p62 and Beclin1 were determined with q-PCR. Flow cytometry was performed to analyze the effect of 50, 100, and 200 μg/mL DMY on cell cycle and apoptosis of HepG2 cells; DNA integrity in the treated cells was examined with cell DNA fragmentation test.@*RESULTS@#DMY treatment and pretreatment obviously inhibited lipid accumulation and reduced the levels of TC, TG, LDL and enzyme activities of AST, ALT and LDH in LO2 cells (P < 0.05). In routinely cultured LO2 cells, DMY significantly promoted the formation of autophagosomes and autophagolysosomes and upregulated the expression of LC3 protein. DMY obviously attenuated high FBS-induced inhibition of autophagosome formation in LO2 cells, up- regulated the mRNA levels of LC3, ATG7, Beclin1 and AMPK, and downregulated p62 and mTOR mRNA levels (P < 0.05 or 0.01). In HepG2 cells, DMY caused obvious cell cycle arrest, inhibited cell proliferation, and induced late apoptosis and DNA fragmentation.@*CONCLUSION@#DMY reduces lipid accumulation in LO2 cells by regulating the AMPK/ mTOR-mediated lipophagy pathway and inhibits the proliferation of HepG2 by causing cell cycle arrest and promoting apoptosis.

The effect and mechanism of epigallocatechol gallate combined with trastuzumab on the proliferation of HER2 overexpressing breast cancer cells / 药学实践杂志

Objective To study the effect and mechanism of epigallocatechol gallate (EGCG) combined with trastuzu-mab on the proliferation of human epidermal growth factor receptor 2 (HER2) overexpressing breast cancer cells. Methods Trastuzumab was expressed and purified. The cell proliferation of HER2 overexpressing breast cancer cells BT474 and SK-BR-3 treated with trastuzumab, EGCG, or trastuzumab plus EGCG was evaluated by CCK8 assay. The effects of EGCG and trastuzumab on the expression of HER2, epidermal growth factor receptor (EGFR), mitogen-activated protein kinase (MAPK), protein kinase B (Akt), and their phosphorylated proteins in BT474 breast cancer cells were detected by Western blot. Results The results of cell proliferation assay indicated that EGCG and trastuzumab, alone or in combination, effectively inhibited the proliferation of BT-474 and SK-BR-3 cells. And within a certain concentration range, EGCG and trastuzumab showed a synergistic proliferation inhibitory effect on HER2 overexpressing breast cancer cells. Consistent with these results, Western blot results showed that trastuzumab and EGCG, alone or in combination significantly reduced the phosphorylation levels of Akt, MAPK, EGFR, and HER2 in BT474 cells. Moreover, the inhibition effect of EGCG plus trastuzumab was significantly more potent than either EGCG or trastuzumab. Conclusion EGCG and trastuzumab could synergistically inhibit the proliferation of HER2 overexpressing breast cancer cells, which may be related to the regulation of Akt and MAPK signaling pathways.

Design, synthesis and anti-cervical cancer activity of novel trifluoromethyl chalcones derivatives / 中国药科大学学报

@#Chalcones are polyphenolic flavonoid substances with various pharmacological effects and low toxicity.In this study, 15 novel trifluoromethyl chalcone derivatives (3a-3o) were designed and synthesized using the chalcone nucleus of natural licorice chalcone as the lead compound skeleton in order to find the candidate drugs with high efficiency and low toxicity against cervical cancer.The structures of the target compounds were confirmed by 1H NMR, 13C NMR and HRMS. The inhibitory activities of compounds 3a-3o, licorice chalcone, cisplatin and Nutlin3a on SiHa, HeLa and C-33A human cervical cancer cells and H8 and HaCaT normal cells were determined by MTT assay, and the structure-activity relationship was analyzed.Transwell and flow cytometry methods were used to assess the target compounds'' ability to inhibit cell migration and invasion, promote apoptosis, and arrest the cell cycle.Molecular docking technology was used to further study the binding characteristics of the target compound with MDM2 protein.The results showed that the compounds had different degrees of inhibitory activity against the three types of cervical cancer cells.Compound 3n showed the strongest activity against HeLa cells (IC50 = 11.69 μmol/L), which was superior to the lead compound, and had lower toxicity against the two normal cells.Compound 3n was found to significantly inhibit the migration and invasion of HeLa cells, induce apoptosis and arrest the cell cycle at G2/M phase.The results of molecular docking showed that the effective binding of compound 3n to MDM2 protein may be one of its anti-tumor mechanisms.This study provides an experimental basis for the screening of new anti-cervical cancer candidate drug from chalcone derivatives.

Aqueous extracts from / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Hepatocellular carcinoma (HCC), which is the most frequent primary liver malignancy, is ranked as the sixth most common cancer and the third leading cause of cancer-related deaths worldwide, with its incidence expected to continue rising. One of the reasons is that most patients are diagnosed at an advanced stage when therapeutic options are ineffective. The development of HCC is attributed to a chronic exposition to either one or a combination of low amounts of different hepatotoxins, such as in hepatitis virus infection, alcohol consumption, aflatoxin from contaminated foods, metabolic factors, and exposure to chemical carcinogens from tobacco smoke (Forner et al., 2018). Integrative studies combining exome sequencing, transcriptome analysis, and the genomic characterization of HCC have shown that these etiological factors may raise the frequency of particular genetic alterations, resulting in intra-tumor heterogeneity that presents a huge challenge for treatment. For example, mutations in the catenin β-1 (CTNNB1) gene (a proto-oncogene in the WNT signaling pathway that encodes the β‍-catenin transcription factor) are strongly associated with alcohol-related HCC, whereas mutations in the telomerase reverse transcriptase (TERT) promoter and tumor protein p53 (TP53) genes are the most commonly observed in hepatitis B virus (HBV)‍-associated HCC (Calderaro et al., 2017; Cancer Genome Atlas Research Network, 2017). The above findings emphasize the molecular diversity of HCC and the associations of different etiologies with distinct mechanisms in HCC progression. Consequently, prevention strategies are still attractive for HCC management.

Effect of Trans-oleic Acid on Proliferation Inhibition and Apoptosis Induction in H9c2 Cardiomyocyte / 中国药学杂志

OBJECTIVE: To investigate the effects and possible mechanism of trans-oleic acid (9t-C18:1) on proliferation inhibition and induction apoptosis in H9c2 cardiomyocyte. METHODS: H9c2 rat cardiomyocytes were cultured in vitro. High, medium and low (600, 300, 150 μmol•L-1) dose of 9t-C18:1 groups and the negative control (NC) group were administered to H9c2 cardiomyocytes. The effect of 9t-C18:1 on cell proliferation was tested using cell counting kit-8 (CCK-8) assay. Morphological changes of cells were observed by AO-EB staining. Intracellular reactive oxygen species (ROS) and apoptosis were detected by flow cytometry. The expression of Bcl-2 and Bax genes was detected by quantitative real time- polymerase chain reaction (QRT-PCR). The expression of Bcl-2 and Bax protein was determined by flow cytometry after immunofluorescence staining. RESULTS: The typical morphological characteristics of apoptosis were observed by fluorescence microscope. The result of CCK-8 assay indicated that 9t-C18:1 have an certainly inhibitory effect on the proliferation of H9c2 cells. ROS level and apoptosis rate were significantly increased. Bcl-2 gene and protein expression were down-regulated, and Bax gene and protein expression were up-regulated, compared with NC group(P<0.05, P<0.01). CONCLUSION: 9t-C18:1 can inhibit the proliferation and induce the apoptosis of H9c2 cardiomyocyte, and its mechanism may be related to promoting the mitochondrial apoptotic pathway.

Effect of Xihuangwan on NLRP3 Inflammatory Bodies and Their Products and Tumor Proliferation of Lung Cancer A549 Bearing Nude Mice in Inflammatory Microenvironment / 中国实验方剂学杂志

Objective:To study the antitumor effect of Xihuangwan on A549 lung cancer nude mice in inflammatory microenvironment, and explore the effect of Xihuangwan on the expressions of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammatory bodies and their products in serum and tumor tissue of A549 lung cancer nude mice. Method:The lung cancer A549 cell model was established in nude mice with lung cancer, and the lung cancer A549 cell model was established in inflammatory microenvironment by adding lipopolysaccharide (LPS) + adenosine triphosphate (ATP) to the culture medium. After modeling, the rats were randomly divided into blank group (equal volume of normal saline), positive drug control group (MCC950 solution, 0.79 g·kg-1), and low, medium, high-dose Xihuangwan groups (0.39, 0.78, 1.95 g·kg-1). The rats were administered orally once a day for 21 days, and then sacrificed. The tumor tissues were stripped to measure the tumor body. The expressions of NLRP3, malondialdehyde（MDA）, interleukin (IL)-1β, IL-18 and NLRP3 were detected by Western blot, and the levels of IL-1β, IL-18 and MDA were detected by enzyme linked immunosorbent assay（ELISA）. Result:Compared with the blank group, the tumor inhibition rates in the positive drug control group and the low, medium and high dose Xihuangwan group were 39.21%, 31.72%, 42.24% and 55.68%. ELISA showed that the high-dose Xihuangwan group could significantly reduce the expressions of MDA, IL-1β and IL-18 in the serum of nude mice (P<0.05). Western blot showed that the high-dose Xihuangwan group could effectively reduce the protein expressions of MDA, IL-1β, IL-18 and NLRP3 in tumor of nude mice (P<0.05), the results of immunohistochemistry showed that the expression rate of NLRP3 in the tumor tissues of nude mice was reduced in the positive drug group and each dose of Xihuangwan group (P<0.05). Conclusion:Xihuangwan can inhibit the growth of tumor tissue of A549 cells bearing lung cancer in nude mice. The mechanism may be that it can inhibit the growth of tumor cells by inhibiting the expression of NLRP3 inflammatory bodies, IL-1β, IL-18, MDA tables, and then inhibiting the inflammatory microenvironment of tumor cells.

Inhibitory effect of isolinderalactone on growth of human breast cancer MCF-7 cells and its mechanism / 中草药

Objective: To study the effects and the mechanism of isolinderalactone on inhibiting the growth of human breast cancer MCF-7 cells. Methods: Human breast cancer MCF-7 cells were cultured in vitro and treated respectively with indicated concentrations of isolinderalactone by cell culture technique. The proliferation rate was detected by MTT method; Flow cytometry and TUNEL assay were used to observe the effects of isolinderalactone on cell cycle, mitochondrial membrane potential and apoptosis in MCF-7 cells; The apoptosis related protein expression levels were determined by Western blotting. Results: Isolinderalactone significantly inhibited the proliferation of MCF-7 cells by inducing cell apoptosis in a time and concentration dependent manner and induced cell cycle arrest at G2/M phases. The mitochondrial membrane potential was depolarized; And isolinderalactone up-regulated the expressions of apoptosis related proteins Bax and cleaved Caspase-3 and down-regulated the expressions of apoptosis related proteins Bcl-2. Conclusion: Isolinderalactone shows obvious anti-cancer activities by inducing cell apoptosis. The mechanism of inducing apoptosis may be associated with the reduction of mitochondrial membrane potential and activation of caspase pathway.

The cell proliferation, cell cycle and apoptosis effects of cycloartane triterpenoid on HCT116 cells / 中国药理学通报

Aim To investigate the antitumor effects of cimigenol ( KY17 ) , a novel cycloartane triterpenoid from Cimicifuga , in human colon cancer cells (HCT116).Methods MTT assay was used to deter-mine the effect of KY17 on the proliferation of mouse embryonic fibroblasts ( MEF) and human colon cancer HCT116 cell line.Flow cytometry was employed to de-tect the effect of KY17 on HCT116 cell cycle.Fluores-cence microscopy and flow cytometry were used to ana-lyze the apoptosis .Western blot was used to detect the expression of apoptotic protein (PARP).q-PCR ana-lyzed the expression of miRNA-34a.Results The IC50 of KY17 in MEF and HCT116 cells was 27.28 μmol· L-1 and 9.31μmol· L-1 , respectively.KY17 induced HCT116 cell cycle arrest in G2/M phase and the apoptotic protein PARP cleavage . In addition, KY17 up-regulated the expression of p 53 protein and miRNA-34a.Conclusions KY17 inhibits the prolifera-tion and the cell cycle is arrested in G 2/M, inducing the apoptosis of HCT116 cells. The mechanism is probably related to miRNA-34a up-regulation and p53 activation .

Effect of 12-Deoxyphorbol-13-Palmitate on Proliferation Inhibition and Apoptosis Induction of HL60 Cells / 中国药学杂志

OBJECTIVE: To investigate the effects of 12-deoxyphorbol-13-palmitate (DP) on the proliferative inhibition and induction apoptosis of human acute myelocytic leukemia cell line HL60. METHODS: HL60 cells were treated with different concentrations of DP. The morphological changes were observed with optical microscope. The effect of DP in cell proliferation was tested using cell counting kit-8(CCK-8) assay, to analyze the cell cloning capacity by methyl cellulose colone-forming experiment. DNA agarose electrophoresis method was used to detect apoptosis. The expression of pro-and anti-apoptotic genes Bcl-2 and Bax were examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: CCK-8 METHODS and methyl cellulose colone-forming experiment indicated DP had an certainly inhibitory effect on HL60 cells(P<0.05, P<0.01). The characteristics of apoptosis was presented under optical microscope. DNA agarose electrophoresis showed the obvious DNA 1adder. In accordance with RT-PCR experiments, compared with the control group(0 mg·L-1), DP could down-regulate the gene expression of Bcl-2 and up-regulate the gene expression of Bax in HL60 cells(P<0.01). CONCLUSION: DP can inhibit the proliferation and induce the apoptosis of cultured HL60 cells in a dose-effect and time-effect relationship in vitro, and its mechanism may be related to down-regulating gene expression of Bcl-2, up-regulating gene expression of Bax, rising Bax/Bcl-2 value.

Mechanism of Photodynamic Therapy Mediated by Hypericin in Inhibiting the Proliferation of Leukemia Cell Lines / 中国药学杂志

OBJECTIVE: To observe the effects of hypericin on proliferation, apoptosis of leukemia cells and its possible mechanism. METHODS: Subcellular localization of hypericin in leukemia cells by fluorescence microscopy. MTT(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) test was adopted to observe the inhibitory effects of hypericin on the proliferation of leukemia cells and calculated its half maximal inhibitory concentration(IC50). The technology of flow cytometry, Annexin-V-FITC/PI double stained method were employed to measure the cell apoptosis of leukemia cells after hypericin treatment. RESULTS: The optimal time for hypericin to entry into cells was 16 h. Hypericin could inhibit the proliferation of leukemia cells in vitro, and the inhibition presented obvious dose-effect relationship(r=0.990 5, P<0.01). The apoptotic ratio of leukemia cells was gradually increased. The ratio of Bax/Bcl-2 was significantly increased, and the apoptotic protein caspase-3, 8, 9 is cleaved and activated. CONCLUSION: Hypericin could inhibit the growth of leukemia cells, induce the apoptosis through induction of caspase dependent apoptosis pathway.

Function of OTUD7B in acute myeloid leukemia cells and its mechanism / 上海交通大学学报（医学版）

Objective: To investigate the biological effect and mechanism of OTUD7B in acute myeloid leukemia (AML) cells. Methods: The expression of OTUD7B in peripheral blood mononuclear cells and bone marrow mononuclear cells of AML patients were detected. The relationship between OTUD7B and survival of AML patients was confirmed by using TCGA database. Mouse model of M2 type AML was established, and the expression of OTUD7B in the bone marrow, spleen and liver of the mice was detected. OTUD7B was overexpressed in AML cell lines HL60 and kasumi1, then the cell viability and cell cycle were measured. The AKT/mTOR pathway proteins were detected after OTUD7B overexpressed and then the cell growth inhibition was detected after overexpression of AKT1. Results: The expression of OTUD7B was lower in primary leukemia cells from all types of AML patients and in the bone marrow, liver and spleen of M2 type AML mice, which was closely related to the survival time of AML patients. OTUD7B overexpression in HL60 and kasumi1 cells significantly inhibited the cell viability and decreased the percentage of S phase cells. OTUD7B significantly inhibited the phosphorylation of AKT and mTOR, and AKT1 overexpression partially reversed the inhibitory effect of OTUD7B on cell growth. Conclusion: OTUD7B expression is low in primary leukemia cells from AML patients and in bone marrow, liver and spleen of the M2 type AML mice. The survival time of patients with low OTUD7B expression is shorter. Overexpression of OTUD7B significantly inhibited the cell viability of HL60 and kasumi1 cells and the entry of cells into S phase. The inhibitory effect of OTUD7B overexpression on AML cells might be related to the inhibition of AKT / mTOR signaling pathway.

A mechanism research of novel inhibitor of PAK1 inducing colorectal cancer DLD-1 apoptosis / 中国药科大学学报

@#PAK1 plays an important role in the development of tumors. It is of great significance to screen and develop new PAK1 inhibitors as targeted drugs for cancer treatment. The traditional PAK1 inhibitor screening method has the problems of high cost and low efficiency. Computer virtual screening can reduce the cost of finding active lead compounds and improve the screening efficiency. In this study, a kind of PAK1 candidate compound was screened by computer assisted virtual screening combined with Z′lyteTM high flux kinase screen. In vitro enzyme activity screening showed that compound 18(K788)had good PAK1 inhibitory activity(inhibition rate was 42. 7%). Furtherly by MTT detection, it was found that K788 had significant PAK1 positive tumor killing activity, which was even better than the positive drug IPA-3. Flow cytometry and Western Blot showed that K788 could activate caspase apoptosis pathway and induce apoptosis of colon cancer cell DLD-1 by inhibiting PAK1 expression and activation. K788 has great potential for clinical development and application, and can be used as a PAK1 target for further research.

Effect of aspirin on the proliferation and apoptosis of human adrenal cortical cancer cells / 中华内分泌代谢杂志

Objective To observe the effect of aspirin on the proliferation inhibition and apoptosis of human adrenal cortical cancer cells, and to explore preliminarily the related mechanisms. Methods The human adrenal cortical cancer cells were cultured in vitro. The experimental group was DEME/F-12 complete medium which contained different concentrations of aspirin (final concentration of 0.125, 0.25, 0.5, 1 mg/ml). The control group was DEME/F-12 complete medium which had no aspirin but 1%anhydrous ethanol instead. After treatment by aspirin at different concentrations for different durations (24, 48, 72 hours), we detected the proliferation inhibition of SW-13 cells and H295R cells by methyl thiazolyl tetrazolium (MTT) method, observed the changes of cell morphology with the inverted microscope; Observed and counted apoptotic cells through Hoechst33258 fluorescent staining, tested the cell apoptosis by flow cytometry, and detected the expression of Bcl-2 and Bax by western blotting. Results The date of MTT showed that after treated by different concentrations of aspirin,the growth inhibition ratio of SW-13 cells and H295R cells were higher than the control group (P＜0.05), and at the same time, the inhibition ratio would increase when the drug concentration increased ( P＜0. 05 ), with a dose-dependent tendency. When the drug concentration was constant, the inhibition ratio increased with the duration of the drug (P＜0.05), which showed a time-dependent tendency. The number of apoptosis cells and the cell apoptosis rate of both SW-13 and H295R which were treated by different concentrations of aspirin for 48 hours were higher than the control group ( P＜0. 01). According to the analysis of grayscale value of western blotting, Aspirin can increase the expression of Bax ( P＜0.05). On the contrary, the expression of bcl-2 in the experimental group was lower than that in the control group (P＜0.05). Conclusion Aspirin may inhibit the growth of human adrenal cortical cancer cell in vitro and induce its apoptosis, and the possible mechanism may be correlated with the up-regulation of the expression of Bax, and down regulation of Bcl-2 expression.

Study on relationship between anti-proliferative effect of resveratrol and BMP7 in human colon cancer cells / 中国药理学通报

Aim To study the relationship between the anti-proliferative effect of resveratrol (Res) and BMP7 on human colon cancer cells and its possible molecular mechanism. Methods The proliferation of HCT116 cells was analyzed with cell proliferation inhibition assay, flow cytometry, Western blot and Annexin V-EGFP staining. CCK-8, PCR and Western blot assay were used to determine the effect of Res on BMP7 in HCT116 cells and the possible molecular mechanism underlying this process. Results Res inhibited the proliferation,arrested cell cycle at S phase and promo-ted apoptosis in HCT116 cells. Res increased the ex-pression of BMP7 mRNA and protein in HCT116 cells. Overexpression of BMP7 enhanced the anti-proliferative effect of Res on HCT116 cells and promoted the Res-induced apoptosis, whereas BMP7-specific antibody significantly attenuated these effects. Res exerted no apparent effect on the phosphorylation of Smad1/5/8, but decreased the phosphorylation of Akt1/2/3 sub-stantially in HCT116 cells. Overexpression of BMP7 enhanced the inhibitory effect of Res on phosphoryla-tion of Akt1/2/3, while BMP7 specific antibody re-duced this effect notably. Res markedly decreased the phosphorylation of PTEN, which could be boosted by BMP7,but attenuated by the BMP7 specific antibody. Conclusions Res can inhibit the proliferation and promote apoptosis of HCT116 cells,and the anti-canc-er activity of Res may be mediated by inactivating PI3K/Akt signaling through up-regulating BMP7 to de-crease the phosphorylation of PTEN partly.

Preparation of Argininate Betulinic Acid and Its Effect of the Proliferation of Triple-negative Human Breast Cancer Cell MDA-MB-231 / 中国药房

OBJECTIVE:To prepare argininate betulinic acid,and to investigate the effect of the proliferation of triple-negative human breast cancer cell MDA-MB-231. METHODS:By using argininate as the solubilization carrier,argininate betulinic acid was prepared by co-grinding equal molar ratio of betulinic acid and argininate. The argininate betulinic acid was characterized with powder X-ray diffractometry,infrared spectroscopy and differential scanning calorimetry. The solubility of betulinic acid and argininate betulinic acid were compared. MTT method was used to assay the effects of 15,30,60,120 μ g/mL betulinic acid, argininate betulinic acid and 5-FU on the proliferation of MDA-MB-231 cell. RESULTS:Prepared argininate betulinic acid was a new phase which was different from the physical mixing of argininate and betulinic acid,among which carboxyl group of betulinic acid and amino group of argininate formed as a salt,and the salt had no obvious melting peak. Betulinic acid was almost insoluble in water. The solubility of betulinic acid in argininate betulinic acid aqueous solution was 50.72 μg/mL. Compared with betulinic acid,the inhibitory rate of argininate betulinic acid on the growth of MDA-MB-231 cell was increased significantly(P<0.05), there was no statistical significance between its effect and 5-FU(P>0.05). CONCLUSIONS:Argininate betulinic acid with good solubility is prepared successfully,and can inhibit the proliferation of MDA-MB-231 cell.

Function of OTUD7B in acute myeloid leukemia cells and its mechanism / 上海交通大学学报（医学版）

Objective·To investigate the biological effect and mechanism of OTUD7B in acute myeloid leukemia (AML) cells.Methods·The expression of OTUD7B in peripheral blood mononuclear cells and bone marrow mononuclear cells of AML patients were detected.The relationship between OTUD7B and survival of AML patients was confirmed by using TCGA database.Mouse model of M2 type AML was established,and the expression of OTUD7B in the bone marrow,spleen and liver of the mice was detected.OTUD7B was overexpressed in AML cell lines HL60 and kasumil,then the cell viability and cell cycle were measured.The AKT/mTOR pathway proteins were detected after OTUD7B overexpressed and then the cell growth inhibition was detected after overexpression ofAKT1.Results·The expression of OTUD7B was lower in primary leukemia cells from all types of AML patients and in the bone marrow,liver and spleen of M2 type AML mice,which was closely related to the survival time of AML patients.OTUD7B overexpression in HL60 and kasumil cells significantly inhibited the cell viability and decreased the percentage of S phase cells.OTUD7B significantly inhibited the phosphorylation of AKT and mTOR,and AKT1 overexpression partially reversed the inhibitory effect of OTUD7B on cell growth.Conclusion·OTUD7B expression is low in primary leukemia ceils from AML patients and in bone marrow,liver and spleen of the M2 type AML mice.The survival time of patients with low OTUD7B expression is shorter.Overexpression of OTUD7B significantly inhibited the cell viability of HL60 and kasumi 1 cells and the entry of cells into S phase.The inhibitory effect of OTUD7B overexpression on AML cells might be related to the inhibition ofAKT / mTOR signaling pathway.

Inhibitory effect of fenbendazole on proliferation of human chronic myelogenous leukemia K562 cells / 中国病理生理杂志

AIM:To investigate the effect of fenbendazole (FBZ) on the proliferation of human chronic myelogenous leukemia (CML) cell line K562.METHODS:The CCK-8 assay was used to detect the effect of FBZ on viability of the K562 cells and normal peripheral blood mononuclear cells (PBMC).The cell growth was measured by the method of Trypan blue exclusion.The cell cycle was analyzed by flow cytometry.The cell cycle-related proteins were detected by Western blot.RESULTS:The growth of K562 was significantly inhibited by FBZ.However, it elicited little cytotoxic effect on PBMC.Furthermore, FBZ induced G2/M phase arrest and mitotic catastrophe in the K562 cells based on the changes of nuclear morphology, DNA content, mitotic marker analysis and the number of polykaryocytes.CONCLUSION:Fenbendazole significantly inhibits the proliferation of K562 cells and induces cell cycle arrest at G2/M phase by the regulation of cell cycle-related proteins.

Biological effects and mechanism of adenanthin on multiple myeloma cells / 上海交通大学学报（医学版）

Objective·To explore the biologic effect and mechanism of adenanthin (Aden) on multiple myeloma (MM) cells. Methods·MM cells, H929 and U266 were treated with various dose of Aden for different time, and the density and viability of MM cells were detected by trypan blue exclusion assay. After H929 and U266 cells were treated with various dose of Aden for 24 hours, cell growth inhibition was examined by CCK8 assay, and cell apoptosis was examined by AnnexinV-APC/PI staining assay. Apoptosis related proteins, NF-κB signaling pathway associated proteins and the NF-κB regulated proteins were detected by Western blotting. The effect of Aden on the thermal stability of IKKβ protein was determined by CETSA assay. Results·Trypan blue exclusion results showed that Aden inhibited cell growth and reduced cell viability in concentration and time dependent manners. U266 was more sensitive than H929 when exposed to the same concentration of Aden. The CCK8 results showed that Aden inhibited the growth of H929 and U266 cells in a concentration dependent manner. Flow cytometry results suggested that Aden induced a low apoptosis rate of MM cells. Moreover, cleavage of caspase3 and PARP were detected in U266 cells but not in H929 cells. CETSA assay indicated that Aden decreased the thermal stability of IKKβ. Expression of p-p65 and p-IκBα proteins decreased in MM cells treated with Aden. Conclusion·Aden significantly inhibits MM cell proliferation by inhibiting NF-κB activation through interacting with IKKβ. Aden has little effect on apoptosis of MM cells.

Influence of Bufalin on the Growth and Proliferation of Human Glioma Cell Line U251 / 中国药师

Objective:To explore the effects of Bufalin on the growth and proliferation of human glioma cells U251. Methods:Methyl thiazolyl tetrazolium(MTT)assay was used to detect the effect of Bufalin on the proliferation of human glioma cells U251. An in-verted microscope was used to observe the changes of cell number,morphology and activity. AnnexinV/ PI was used to measure the in-duction of cell apoptosis caused by Bufalin. Results:Bufalin at different concentrations(0. 001 - 10. 0μmol·L - 1 )inhibited the pro-liferation of U251 cells in a dose and time-dependent manner. Compared with that of the control group,the apoptosis rate of Bufalin group was increased significantly(P < 0. 01). Conclusion:Bufalin can inhibit the growth and proliferation of U251 cells in a dose and time-dependent manner,and induce the apoptosis of U251 cells.

Preparation and Tumor Inhibition Effect of Transferrin Modified Paclitaxel-loaded Liposome / 中国药房

OBJECTIVE:To prepare transferrin modified paclitaxel-loaded liposome(TF-PTX-LP),and to study the tumor in-hibition effect. METHODS:TF-PTX-LP was prepared by thin-film method,and morphology of TF-PTX-LP was observed. Qualita-tive and quantitative investigation were used to value the uptake efficiency of TF-LP and LP by HepG2 cells. The proliferation inhi-bition rate of HepG2 cells was investigated after treated with PTX,PTX-LP and TF-PTX-LP for 24,48 and 72 h. Tumor spheres were prepared by using HepG2 cells. Effects of normal saline,PTX,PTX-LP and TF-PTX-LP on the volume of tumor spheres were investigated after 0,1,2,4,5,6 and 7 d treatment. HepG2 tumor-bearing nude mice model was induced. Inhibitory effects of normal saline,PTX,PTX-LP and TF-PTX-LP(8.5 mg/kg by PTX)on transplantable tumor of tumor-bearing nude mice were in-vestigated. RESULTS:TF-PTX-LP showed uniform spherical shape,with particle size of 100-120 nm. The fluorescence intensity of HepG2 cells treated with TF-LP was stronger than that treated with LP(P<0.01). Compared with PTX and PTX-LP,TF-PTX-LP showed higher proliferation inhibition rate(P<0.01). Compared with normal saline,PTX and PTX-LP,tumor spheres were small-er in volume after treated with TF-PTX-LP,and inhibition rate of tumor was higher in tumor-bearing nude mice;there were statisti-cal significance after treated for 6,7 d(P<0.01). The proliferation inhibition rate and tumor spheres volume changed in time-de-pendent manner. CONCLUSIONS:TF-PTX-LP which owns good tumor inhibition effect is prepared successfully.