Proliferation Inhibitory Effect Research Articles

Prenatal dexamethasone exposure caused fetal rats liver dysplasia by inhibiting autophagy-mediated cell proliferation

As a synthetic glucocorticoid, dexamethasone has been widely used in the clinical treatment of premature birth and related pregnant diseases, but its clinical use is still controversial due to developmental toxicity. This study aimed to confirm the proliferation inhibitory effect of pregnant dexamethasone exposure (PDE) on fetal liver development and elucidate its molecular mechanism. In vitro studies, we found that dexamethasone inhibited hepatocyte proliferation through autophagy activated by glucocorticoid receptor (GR)-forkhead protein O1 (FOXO1) pathway. Subsequently, in vivo, we confirmed in a PDE rat model that male fetal liver proliferation was inhibited, and the expression of the GR-FOXO1 pathway and autophagy were increased. Taken together, PDE induces autophagy by activating the GR-FOXO1 pathway, which leads to fetal liver proliferation inhibition and dysplasia in offspring rats. This study confirmed that dexamethasone activates cell autophagy in utero through the GR-FOXO1 pathway, thereby inhibiting hepatocyte proliferation and liver development, which provides theoretical basis for understanding the developmental toxicity of dexamethasone and guiding the rational clinical use.

Experience with regorafenib in the treatment of hepatocellular carcinoma.

Regorafenib is a diphenylurea oral multikinase inhibitor, structurally comparable to sorafenib, which targets a variety of kinases implicated in angiogenic and tumor growth-promoting pathways. Regorafenib was the first agent to positively show significant survival advantage as a second-line therapy in patients with unresectable hepatocellular carcinoma (HCC) who had previously failed first-line treatment with sorafenib. Recent evidence has shown that its antitumor efficacy is due to a comprehensive spectrum of tumor neo-angiogenesis and proliferation inhibition and immunomodulatory effects on the tumor microenvironment, which plays a crucial role in tumor development. This review addresses the rationale and supporting evidence for regorafenib’s efficacy in HCC that led to regorafenib’s approval as a second-line therapy. In addition, we review proof from clinical practice studies that validate the RESORCE trial results. We discuss regorafenib’s potential role in the newly emerging therapeutic strategy based on combination with immune checkpoint blockade and its possible extensibility to patient categories not enrolled in the registrative study.

Identification of poly(ADP-ribose)polymerase 1 and 2 (PARP1/2) as targets of andrographolide using an integrated chemical biology approach.

Here, we describe the identification of PARP1/2 as direct binding proteins of andrographolide (Andro) using protein microarray, surface plasmon resonance (SPR), and enzyme activity assays. We then evaluated the proliferation inhibition, apoptosis, and cell migration effects of Andro on the MDA-MB-436 cell line in vitro. The final biological evaluation confirmed that Andro was a highly effective single agent in the MDA-MB-436 xenograft model and had a low hERG-mediated cardiac toxicity. Therefore, Andro represents the first natural product, non-amide member of a novel nanomolar-potency PARP1/2 inhibitor family.

Novel poly (ADP-ribose) polymerases inhibitor DHC-1 exhibits in vitro and in vivo anticancer activity on BRCA-deficient pancreatic cancer cells

Poly (ADP-ribose) polymerases (PARPs) play a key role in DNA repair. In this study we designed a novel small-molecular compound, (E)-2-(2,3-dibromo-4,5-dimethoxybenzylidene)hydrazine-1-carbothioamide (DHC-1), which was a potent and selective PARP-1 inhibitor. DHC-1 selectively inhibited PARP-1 activity with an IC50 value of 41.12 ± 13.28 nM. Cytotoxicity results showed that DHC-1 selectively inhibited the proliferation of BRCA1-deficient breast cancer HCC-1937 and BRCA2-deficient pancreatic cancer Capan-1 cells. Mechanism studies found that DHC-1 stabilized PARP-1–DNA complexes and inhibited PAR formation in BRCA2−/− Capan-1 cells. Further experiments found that DHC-1 induced DNA double-strand damage in BRCA2−/− Capan-1 cells, which was demonstrated by accumulation of γ-H2AX foci. Flow cytometry experiments revealed that DHC-1 induced G2/M phase arrest and activate mitochondrial-induced apoptotic pathways. Interestingly, we also found that DHC-1 enhanced cell proliferation inhibitory effect of oxaliplatin (OXA). The further in vivo nude mouse studies showed that DHC-1 inhibited the growth of Capan-1 xenografts and showed a similar mechanism to that in vitro. Collectively, our results demonstrate that DHC-1 may be an excellent candidate for treatment of BRCA-deficient pancreatic cancers.

Chloroform extract of Citrus unshiu Markovich peel induces apoptosis and inhibits stemness in HeLa human cervical cancer cells

Cervical cancer is the second most common cancer among women worldwide. However, chemotherapies for this cancer often cause many side effects and chemoresistance. Citrus unshiu Markovich peel (CECU) has been used as a traditional medicine for the treatment of various diseases in East Asia. Recently, the anticancer activities and mechanisms of action of CECU extract have been reported in a number of different cancer cell types, but no study has evaluated the therapeutic effect of this natural product on cervical cancer cells. In the current study, the anticancer activity and the underlying molecular mechanism of the chloroform extract of CECU was investigated on HeLa human cervical cancer cells. The results showed that CECU effectively inhibited the proliferation and migration of HeLa cells. Treatment of cells with CECU led to cell cycle arrest at the G2/M phase and activation of extrinsic and intrinsic apoptotic pathways. Furthermore, the proliferation inhibitory effect of CECU was due to the inactivation of AKT and ERK signaling, upregulation of p53 and p21, and downregulation of cyclin B1 and cyclin D1, but not reactive oxygen species (ROS) generation. Furthermore, CECU inhibited the stem-like features of HeLa cells by downregulating key cancer stemness biomarkers. Therefore, CECU may be an effective complementary and alternative medicine for the prevention and treatment of cervical cancer.

Fe3O4 Nanoparticles Capped with PEG Induce Apoptosis in Breast Cancer AMJ13 Cells Via Mitochondrial Damage and Reduction of NF-κB Translocation

In the current study, polyethylene glycol (PEG) was employed to cap super magnetite nanoparticles (Fe3O4 NPs) through hydrothermal preparation. The main goal of this study is to investigate the influence of physical incorporation of polyethylene glycol (PEG) loaded Fe3O4. The anticancer potentials of these particles were studied against breast cancer cell line (AMJ13). Syntheses bare Fe3O4, and Fe3O4-PEG were confirmed by TEM, SEM, and FTIR. The size of Fe3O4 nanoparticles range of 9–20 and 5–12 nm for the Fe3O4–PEG nanoparticles which exerted superparamagnetic properties as well as elevated saturation level of magnetization at ambient conditions. The MTT test was employed to detect the ability of the bare Fe3O4 and Fe3O4-PEG on the proliferative of AMJ13 cells. IC50 values was 37.33 µg mL− 1 for bare Fe3O4 and 18.23 µg mL−1 for Fe3O4-PEG. AMJ13 Cells exposed to bare Fe3O4, and Fe3O4-PEG NPs demonstrated a significant cell death, which increased with PEG, loaded Fe3O4 NPs. The capability of Fe3O4-PEG to induce cellular apoptosis was tested using DAPI, Acridine orange/ Ethedium bromide stains, flow cytometry, with the assays of mitochondrial membrane potential (MMP), and the production of reactive oxygen species (ROS). RT-PCR, and immunofluorescence were performed to measure expression levels of Bax and Bcl-2 proteins. The toxicity of bare Fe3O4 and Fe3O4-PEG nanoparticles using animal model were investigated. Animal’s body weight, liver and kidney function enzymes, and histological alterations for liver, kidney, and lungs were addressed. The findings demonstrated that nanoparticles were biocompatible with liver and kidney function enzymes and no significant alterations were recorded in the liver, kidney and the lungs. Both nanoparticles revealed a proliferation inhibitory effect on AMJ13 cells, resulting in apoptosis as a novel pathway that involve the mitochondrial damage and NF-kB. Taken together the present data suggest that bare Fe3O4 and Fe3O4-PEG could be promising therapy protocol for cancer cells.

Tumor cell membrane-derived nano-Trojan horses encapsulating phototherapy and chemotherapy are accepted by homologous tumor cells

Tumor cell membrane-derived nanostructures targeting homologous tumors are promising biomimetic drugs. Herein, curcumin (Cur) and chlorin e6 (Ce6) were co-loaded into PLGA nanoparticles (NPs), and then the NPs were coated with MCF-7 cell membranes (MCNPs). Cell membrane coating sharply increased the uptake of MCNPs by homologous cells, as compared to that with naked NPs. The NPs co-loaded with Cur and Ce6 (Cur/Ce6-NPs) showed a stronger proliferation-inhibitory effect on MCF-7 cells than the NP groups loaded with Cur and Ce6 alone. Cytotoxicity and apoptosis rates of MCF-7 cells in the Cur/Ce6-MCNPs group were significantly higher than those in the uncoated Cur/Ce6-NPs group. Both Cur/Ce6-NPs and Cur/Ce6-MCNPs significantly inhibited the migration of MCF-7cells, although Cur/Ce6-MCNPs showed a stronger effect. Compared to that of Cur/Ce6-NPs, the elimination of Cur/Ce6-MCNPs was both decreased and retarded, prolonging their in vivo systemic circulation and resulting in improved bioavailability. After intravenous administration for 24 h, the fluorescence intensity of drugs in the liver and spleen of the Cur/Ce6-MCNPs group was significantly weaker than that in the Cur/Ce6-NPs group, but that in tumor tissue was enhanced. Further, Cur/Ce6-MCNPs treatment achieved significantly better tumor-suppressive effects in vivo than Cur/Ce6-NPs, resulting in smaller tumor weights, increased apoptosis rates, and the down regulation of Ki67 protein in the tumor tissue. Thus, the tumor cell membrane-camouflaged nanocomposites have potential for homologous tumor-targeted therapy. Furthermore, photodynamic therapy combined with chemotherapy has promising future prospects.

Preparation, characterization and in vitro antitumor activity evaluation of hyaluronic acid-alendronate-methotrexate nanoparticles

As an anti-metabolic drug, methotrexate (MTX) plays an important role in the treatment of various malignant tumors. However, several side effects such as low selectivity and high toxic of MTX limited its further applications. With aims to increase its accumulation in the tumor sites and reduce the toxicity of normal tissue nonspecific uptake, a self-assembled hyaluronic acid-alendronate-methotrexate nanoparticle (HA-ALN-MTX NPs) with a dual-tumor-targeted drug loaded system was designed and synthesized with an average particle size of 265.6 ± 13.3 nm. The advantage of this nanosystem is that the anticancer drug MTX can be used as a tumor-targeted ligand for folate acid receptors (FA), and hyaluronic acid (HA) can be used as another tumor targeted ligand for CD44 receptors. In vitro experiments confirmed that HA-ALN-MTX NPs has lower toxic effect on normal tissue cells HUVECs and has relatively high proliferation inhibition effect on tumor cells A549. Moreover, the inhibition effect could be adjusted by altering the dose of given drugs. All these results revealed that the prepared HA-ALN-MTX NPs could be selectively taken up by tumor cells by FA and CD44 receptor-mediated endocytosis. Therefore, self-assembled HA-ALN-MTX NPs targeted by these FA/CD44 receptors for anticancer drugs could act as effective antitumor drugs.

Beyond Assurance and Coercion: US Alliances and the Psychology of Nuclear Reversal

This article examines the proliferation-inhibiting effect of US alliances. Existing explanations for nuclear reversals of US allies have focused either on the assurance that alliances provide or on US threats of abandonment. However, neither model can account for the fact that allied leaders disagreed over the reversal decision. Also, whether an ally agreed to or rejected nuclear restraint depended on which policymakers carried the day as much as on external factors. To explain why some policymakers accept and others refuse nuclear reversal, I draw on a psychological aspect of US alliances: the social pressure inherent in demands by the United States as an ally holding a superior international status. New evidence from Germany and South Korea shows only policymakers who acknowledge this higher rank of the United States, and hence view their own nation as inferior, respond to this social pressure by obeying the US demand for a nuclear reversal.

Preparation and Evaluation of Collagen-Based Patches as Curcumin Carriers.

Patients with psoriasis are dissatisfied with the standard pharmacological treatments, whether systemic or topical, with many of them showing interest in complementary and alternative medicine. Curcumin (Cur), a natural polyphenol derived from turmeric, has recently gained attention for skin-related diseases because of its proven anti-inflammatory action. However, topical treatment with Cur would be inadequate because of its hydrophobicity, instability, and low bioavailability. In addition, hyperkeratosis and lack of moisture in psoriatic skin result in low penetration that would prevent actives from permeating the stratum corneum. In this work, a polymer-based formulation of Cur for the topical treatment of psoriasis is reported. To improve the physicochemical stability of Cur, it was first encapsulated in chitosan nanoparticles. The Cur-loaded nanoparticles were incorporated in a hydrophilic, biocompatible collagen-based patch. The nanoparticle-containing porous collagen patches were then chemically cross-linked. Morphology, chemical interactions, swelling ratio, enzymatic hydrolysis, and Cur release from the patches were evaluated. All patches showed excellent swelling ratio, up to ~1500%, and after cross-linking, the pore size decreased, and their hydrolysis rates decelerated. The in vitro release of Cur was sustained with an initial burst release, reaching 55% after 24 h. Cur within the scaffolds imparted a proliferation inhibitory effect on psoriatic human keratinocytes in vitro.

Akebia saponin E, as a novel PIKfyve inhibitor, induces lysosome-associated cytoplasmic vacuolation to inhibit proliferation of hepatocellular carcinoma cells

Ethnopharmacological relevanceHepatocellular carcinoma (HCC) is an aggressive malignancy with increasing mortality in China. Screening and identifying effective anticancer compounds from active traditional Chinese herbs for HCC are in demand. Akebia trifoliata (Thunb) Koidz, with pharmacological anti-HCC activities in clinical, has been shown in previous research. In the present research, we elucidated a potential anticancer effect of Akebia saponin E (ASE), which is isolated from the immature seeds of Akebia trifoliata (Thunb.) Koidz, and revealed that ASE could induce severe expanded vacuoles in HCC cells. But the potential mechanism of vacuole-formation and the anti-HCC effects by ASE remain uncover. Aim of this studyTo elucidate the potential mechanism of vacuole-formation and the proliferation inhibition effects by ASE in HCC cell lines. Materials and methodsMTT assay, colony formation assay and flow cytometry were performed to detect cell viability. Immunofluorescence analysis was used to examine the biomarkers of endomembrane. Cells were infected with tandem mRFP-GFP-LC3 lentivirus to assess autophagy flux. RNA-seq was conducted to analyze the genome-wide transcriptional between treatment cell groups. In vitro PIKfyve kinase assay is detected by the ADP-GloTM Kinase Assay Kit. ResultsASE could inhibit the proliferation of HCC with severe expanded vacuoles in vitro, and could significantly reduce the size and weight of xenograft tumor in vivo. Further, the vacuoles induced by ASE were aberrant enlarged lysosomes instead of autophagosome or autolysosomes. With cytoplasmic vacuolation, ASE induced a mTOR-independent TFEB activation for lysosomal biogenesis and a decrement of cholesterol levels in HCC cells. Furthermore, ASE could reduce the activity of PIKfyve (phosphoinositide kinase containing a FYVE-type finger), causing aberrant lysosomal biogenesis and cholesterol dyshomeostasis which triggered the expanded vacuole formation. ConclusionASE can prospectively inhibit the kinase activity of PIKfyve to induce lysosome-associated cytoplasmic vacuolation, and may be utilized as an alternative candidate to treat human HCC.

Effect of Huaier Aqueous Extract Combined with Routine Chemo-therapeutic Drugs on Human Acute Lymphoblastic Leukemia Cells Nalm-6 and Sup-B15

To investigate the proliferation inhibition and pro-apoptotic effect of Huaier aqueous extract combined with routine chemotherapeutic drugs including Vincristine (VCR), Daunorubicin (DNR), L-aspartase (L-Asp) on human acute lymphoblastic leukemia cell lines Nalm-6 and Sup-B15. Nalm-6 and Sup-B15 cell lines were treated with different concentrations of Huaier aqueous extract and chemotherapeutics including VCR, DNR, L-Asp alone or in combination for 48 h, and the growth inhibitory effect and IC50 values (the half maximal inhibitory concentration) were detected by CCK-8. Jin's formula was used to estimated the synergistic effect of these combinations. Apoptosis rates of Nalm-6 and Sup-B15 cells and expression of apoptosis-related proteins BAX, BCL-2, cleaved Caspase-3 were determined by flow cytometry and Western blot respectivcly. Huaier aqueous extract, VCR, DNR and L-Asp had inhibition effect on Nalm-6 and Sup-B15 cell lines. The inhibition rate of Huaier aqueous extract combined with VCR, DNR and L-Asp were all higher than those of each dug alone (P＜0.05) and the combination index (q) was between 0.85 and 1.15 or greater than 1.15. The two kinds of drugs showed had additive or synergistic effects. The results of flow cytometry showed that the cell apoptosis rates in combined treatment group were higher than those of each drug alone (P＜0.05). The results of Western blot revealed that Huaier aqueous extract and VCR all decreased protein expression of BCL-2 (P＜0.05) and increase protein expression of BAX (P＜0.05) and cleaved Caspase-3 (P＜0.05) in Nalm-6 and Sup-B15 cells. Compared with Huaier aqueous extract or VCR alone, the effect of two drug combination were more significant. DNR down-regulated protein expression of BCL-2 (P＜0.05) and up-regulated cleaved Caspase-3 (P＜0.05). However, it had no effect on the expression of BAX in Nalm-6 and Sup-B15 cells. When it was combined with Huaier aqueous extract, the expression of cleaved Caspase-3 and BCL-2 showed more significant changes. The expression of BAX in combined treated group did not show significant difference, compared with group treated with Huaier aqueous extract in Nalm-6 and Sup-B15 cells. L-Asp did not show significant effect on the three apoptosis-related proteins and there was no significant difference between the combination group and the Huaier aqueous extract group. the combination of Huaier aqueous extract and VCR, DNR, L-Asp shows additive or synergistic effects on human acute lymphoblastic leukemia cell lines Nalm-6 and Sup-B15.

Proliferation-Inhibitory and Apoptosis-Inducing Effects of Targeting MTH1 on Multiple Myeloma Cells

To detect the expression of MutT homolog 1 (MTH1) in CD138-negative cells (CD138-) and CD138-positive cells (CD138+) cells of the patients with multiple myeloma (MM), and to explore the effect of MTH1 inhibitor TH588 on cell morphology, proliferation and apoptosis of MM cell U266. CD138- and CD138+ cells of MM patients were isolated, and RNA was extracted. The expression of NUDT family was detected by Q-PCR. MM cell line U266 was used to observe the effect of IL-6 on MTH1 expression. Using fluorescence microscopy to observe the morphological changes of U266 after treatment by TH588 for 48 hours. DAPI staining was used to investigate the nuclear change. Luciferase was used to detect the effect of TH588 on U266 cell proliferation. After treatment with TH588 for 48 h, the change of apoptosis in MM U266 cells was detected by flow cytometer. In some MM patients, the expression of MTH1, NUDT2, NUDT5 and NUDT21 in CD138+ cells was higher than that in CD138- (P<0.05). The luciferase report showed that after treatment of U266 cells with TH588, the fluorescence intensity was significantly lower than that of the control group, and the fluorescence intensity decreased with the increase of drug concentration (r=-0.91). IL-6 could increase the expression of MTH1. Fluorescence microscopy showed that after TH588 treatment of U266 cells for 48 hours, the cells appeared shrinking, smaller, and irregular in shape. After DAPI staining, the TH588-treated cells showed apoptosis characteristics, such as nuclear shrinkage, uneven staining, petal, and radial shape. Flow cytometry showed that the proportion of U266 viable cells decreased significantly after treatment with TH588 for 48 h (P<0.05). MTH1 highly expresses in CD138+ cells of some MM patients. MTH1 expression can increase in U266 cells treated by IL-6. The MTH1 inhibitor TH588 possesses proliferation-inhibitory effect and apoptosis-inducing effect on MM cell U266.

Anti-TMV activity and mode of action of three alkaloids isolated from Chelidonium majus.

Plant viral diseases are difficult to control and have caused serious damage to the agricultural industry. Recently, botanical biopesticides characterized by environment friendly, safe to non-target organism and not as susceptible to produce drug resistance, have exhibited great potential to be developed as antiviral agents. To screen the natural products with antiviral effect, three alkaloids possessed anti-tobacco mosaic virus (TMV) activity were isolated from Chelidonium majus and the modes of action were investigated. The anti-TMV effect of crude extracts at 10 mg mL-1 was 51.73%. Bioassay-guided fractionation and isolation of the compounds with anti-TMV activity were performed on the methanol extract of C. majus yielding three bioactive alkaloids namely: chelerythrine (1), chelidonine (2), and sanguinarine (3). The results of bioassay showed that chelerythrine exhibited great inactivation, proliferation inhibition and protection effects against TMV at 0.5 mg mL-1 with the efficiency of 72.67%, 77.52% and 59.34%, respectively. Chelidonine at 0.1 mg mL-1 can provide 54.90% and 64.45% inhibitions on TMV through inducing resistance in two kinds of tobacco. Sanguinarine showed a weaker protection for resisting TMV in comparison to chelerythrine and chelidonine. Chelerythrine and chelidonine displayed significant inhibitions on TMV with different modes of action. These results provided important evidence that the extracts in C. majus might be a potential source of new drugs in controlling virus disease agriculturally.

Effects and Mechanisms of Acylated Flavonoid on Cancer Chemopreventive Activity

Many studies have shown that flavonoids have various physiological activities such as anti-inflammatory, antibacterial, anti-cancer, and hepatoprotective activities. Meanwhile, its hydrophilic nature has led to low absorption into the body and the limitations of its bioactivity. Many acylated flavonoids and their bioactivities, including those obtained as natural compounds and those synthesized biochemically or chemically, have been reported to improve their bioavailability. This review focuses on the recent developments of the cancer chemopreventive activity of acylated flavonoids. Novel acylated flavonoids have been identified from nature since 2015, and many of them have shown high cytoprotective and anti-inflammatory effects by acylation of glycosyl residues. A large number of acylated flavonoids have been synthesized by enzyme-catalyzed and chemical methods, and their bioactivity has been evaluated. The introduction of acyl groups in the sugar residues showed higher antioxidant activity with increased lipophilicity and improved anti-inflammatory and cytoprotective effects and the ability of apoptosis induction in cancer cells. In contrast, introducing acyl groups into the hydroxy groups of aglycones resulted in less antioxidant activity but improved cancer proliferation inhibition and anti-inflammatory effects. Acylation of flavonoids tends to increase their bioactivity over the parent compound. The increased bioactivity by acylation can be attributed to increased lipophilicity and improved membrane permeability. Acylated flavonoids are one of a group of promising compounds that have cancer chemopreventive effects. However, the study of ADME (Absorption, Distribution, Metabolism, Excretion) will be essential to elucidate the relationship between acylation of flavonoids and the improvement of their bioactivity.

MicroRNA-221/222 Inhibits the Radiation-Induced Invasiveness and Promotes the Radiosensitivity of Malignant Meningioma Cells.

The controversy of adjuvant radiotherapy of meningiomas is at least partially due to the insufficient understanding on meningioma cells' response to irradiation and the shortage of radiosensitivity-promotion methods. MicroRNA-221 and microRNA-222 were identified as critical regulators of radiosensitivity in several other tumors. However, their effect in meningiomas has yet to be confirmed. Therefore, the malignant meningioma IOMM-Lee cells were adopted, transfected with microRNA-221/222 mimics or inhibitors, and irradiated with different dosages. The effects of radiation and microRNA-221/222 were then assessed in vitro and in vivo. Radiation dose increases and microRNA-221/222 downregulation synergistically inhibited cell proliferation and colony formation, prevented xenograft tumor progression, and promoted apoptosis, but antagonistically regulated cell invasiveness. Pairwise comparisons revealed that only high-dose radiations (6 and 8 Gy) can significantly promote cell invasiveness in comparison with unirradiated counterparts. Further comparisons exhibited that downregulating the microRNA-221/222 expression can reverse this radiation-induced cell invasiveness to a level of untransfected and unirradiated cells only if cells were irradiated with no more than 6 Gy. In addition, this approach can promote IOMM-Lee's radiosensitivity. Meanwhile, we also detected that the dose rate of irradiation affects cell cycle distribution and cell apoptosis of IOMM-Lee. A high dose rate irradiation induces G0/G1 cell cycle arrest and apoptosis-promoting effect. Therefore, for malignant meningiomas, high-dose irradiation can facilitate cell invasiveness significantly. Downregulating the microRNA-221/222 level can reverse the radiation-induced cell invasiveness while enhancing the apoptosis-promoting and proliferation-inhibiting effects of radiation and promoting cell radiosensitivity.

MiR-199a-3p suppresses progression of esophageal squamous cell carcinoma through inhibiting mTOR/p70S6K pathway.

Dysregulation of microRNA contributes to multiple tumorigenic processes. Although downregulation of miR-199a-3p has been shown in many cancers, its effects on esophageal squamous cell carcinoma (ESCC) and the regulatory mechanism are still obscure. Here, we aim to evaluate the biological function and underlying mechanisms of miR-199a-3p in ESCC as well as its value to clinical treatment of ESCC. We first analyzed expression of miR-199a-3p in esophageal cancer by bioinformatic analysis and found that there were different opinions about expression of miR-199a-3p in esophageal cancer, and the following qRT-PCR assay demonstrated which was markedly downregulated in ESCC cells. Next, we increased the expression of miR-199a-3p in ESCC cells using miR-199a-3p mimics and demonstrated that overexpression of miR-199a-3p significantly inhibited cell proliferation, migration and invasion, as well as induced cell cycle retard and promoted apoptosis in ESCC. Furthermore, we explored the functional targets of miR-199a-3p and identified that overexpression of miR-199a-3p inhibited mTOR/p70S6K pathway, but stimulated PI3K/Akt pathway. Finally, we demonstrated that overexpression of miR-199a-3p enhanced proliferation-inhibiting effects of MK2206, an inhibitor of Akt, to ESCC cells, which might be related that MK2206 eliminated the activation of miR-199a-3p to p-Akt. These findings discover that miR-199a-3p might participate in the carcinogenesis process of ESCC, which provides a new insight for treatment of ESCC.

IN VITRO AND IN SILICO STUDIES ON EVALUATING EREMANTHIN AS POTENTIAL THERAPEUTIC DRUG AGAINST BREAST CANCER

Objective: The present study was aimed to evaluate the anticancer property of eremanthin isolated from Costus speciosus against breast cancer using in vitro and in silico approaches and thereby to develop eremanthin as a typical phytotherapeutic drug against cancer.&#x0D; Methods: The presence of specific biologically active extract was confirmed under GC–MS/MS (gas chromatography–mass spectrometry) analysis. The cell proliferation inhibitory effect of the eremanthin was confirmed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) and LDH (lactate dehydrogenase) assay. In silico studies were performed to predict the targeted interaction of eremanthin with cancer proteins.&#x0D; Results: The GC–MS/MS analysis confirmed the presence of eremanthin with peak value of RA: 20.03. The MTT and LDH assays revealed the antiproliferative activity of eremanthin on MCF-7 and MDA-MB-231 breast cancer cell lines. The results provide stable interaction between eremanthin and cancer target proteins.&#x0D; Conclusion: Thus, the compound can be used as an effective herbal therapeutic molecule to treat cancer with further explorations.

MiR-199a-5p Affects Sensitivity of Acute Myeloid Leukemia to Adriamycin by Targeting DRAM1

To compare the expression of miR-199a-5p between ADM-resistant AML cell (K562/ADM)and ADM-sensitive AML cell (K562), and to investigate the effect of miR-199a-5p on regulating AML drug resistance as well as its molecular mechanism. MTT method was used to detect the proliferation inhibition effect of ADM on K562 and K562/ADM cells, the IC50 was calculated. miR-199a-5p expression in cell lines (K562 and K562/ADM) and bone marrow sample (refractory/relapsed AML patients and complete remission AML patients) was detected by RT-qPCR. K562/ADM and K562 cells were transfected by miR-199a-5p mimic and miR-199a-5p inhibitor respectively to ensure that miR-199a-5p expression in K562/ADM cells was increased and that in K562 cells was decreased. Then proliferation inhibition effect of ADM on both cells was detected by CCK-8 and mRNA and protein DRAM1 expression in both cells was measured by real time RT-PCR and Western blot respectively. Dual luciferase reporter assay was used to detect wether there were direct binding sites between miR-199a-5p and DRAM1 3' UTR. CCK-8 was used to measure the proliferation inhibition effect of ADM on K562/ADM cells when DRAM1 was downregulated by siRNA. The IC50 of ADM for K562/ADM and K562 cells was 146.14±0.079 and 3.08±0.056 μg/ml respectively. As compared with patients in complete remission group, MiR-199a-5p expression in refractory/ relapsed AML patients significantly decreased, and the MiR-199a-5p expression in K562/ADM cells was also dramatically downregulated, compared with K562 cells (P＜0.05). When the expression of miR-199a-5p was upregulated in K562/ADM cells, the proliferation inhibition effect of ADM on cells elevated and both DRAM1 mRNA and protein expressions decreased. Conversely, when miR-199a-5p expression was downregulated in K562 cells, the proliferation inhibition effect of ADM on cells obviously reduced and both DRAM1 mRNA and protein expression increased (P＜0.05). Dual luciferase reporter Assay showed a direct interaction between miR-199a-5p and its binding site within DRAM1 mRNA. Both DRAM1 mRNA and protein expression in K562/ADM were markedly higher than those in K562 cells (P＜0.05). The ADM chemosensitivity of K562/ADM cells was improved significantly when DRAM1 expression was downregulated (P＜0.05). miR-199a-5p is downregulated in chemoresistant AML cells. miR-199a-5p expression plays an important role in regulating the sensitivity of AML cells to ADM treatment. DRAM1 is a functional target gene for miR-199a-5p modulating AML chemoresistance.

A Multitarget Gold(I) Complex Induces Cytotoxicity Related to Aneuploidy in HCT-116 Colorectal Carcinoma Cells.

A novel alkynyl phosphane gold(I) complex (trimethylphosphane)(3‐(1,3‐dimethylxanthine‐7‐yl)prop‐1‐yn‐1‐yl)gold(I) 1 displayed mutiple biological activites including selective proliferation inhibitory, anti‐metastatic, and anti‐angiogenic effects. The complex also induced effects related to aneuploidy in HCT‐116 colon carcinoma cells, which might be mainly ascribed to the dysfunction of mitochondrial bioenergetics and downregulation of glycolysis. Induction of aneuploidy beyond a critical level can provide an effective strategy to target cancer, in particular colorectal tumours with a low tolerance of aneuploidy, and could be of relevance for 1 and other metallodrugs.