Andrographolide Inhibits Proliferation of Colon Cancer SW-480 Cells via Downregulating Notch Signaling Pathway

Andrographolide Inhibits Proliferation of Colon Cancer SW-480 Cells via Downregulating Notch Signaling Pathway

Background: Recently, Notch signaling pathway has gained attention as a potential therapeutic target for chemotherapeutic intervention. However, the efficacy of previously known inhibitors of Notch in colon cancer remains unclear. The purpose of this study was to determine the effect of andrographolide in aberrantly activated Notch signaling in SW-480 cells in vitro.


Methods: The potential cytostatic andrografolida on SW-480 cells was evaluated by 3- (4,5-dimethylthiazol-2-yl) -2, 5-diphenyl tetrazolium bromide (MTT) assay, assessment of morphology and colony formation test. FITC apoptosis activity was evaluated by Annexin V assay, 4 ‘, 6-diamidino-2-phenylindole (DAPI), Hoechst, Rhodamine 123 and Mito Tracker staining CMXRos. Scratching assay for assessment of potential migration. 7’-Dichlorodihydrofluorescein Diacetate (DCFH-DA) staining was used to evaluate Reactive Oxygen Species (ROS) generation. mRNA relative expression of Bax, Bcl-2, NOTCH 1 and jagged 1 was estimated by real-time quantitative reverse transcription PCR (qRT-PCR). cell cycle phase distribution was evaluated Annexin V-FITC / PI staining.


Results: MTT assay showed a dose and time dependent cytoxicity of andrographolide on SW-480 cells. It also inhibits migratory and colony forming potential of SW-480 cells. In addition, andrografolida also showed impaired mitochondrial membrane potential and induced apoptosis through nuclear condensation. Flow cytometric evaluation showed improved early and late andrographolide cells induced apoptosis and upregulation of proapoptotic (Bax and Bad) and downregulation of Bcl-2 antiapoptotic in SW-480 treated cells. Andrographolide plus intracellular ROS and induced G0 / G1 phase cell cycle arrest in SW480 colon cancer cells. Furthermore, andrografolida suppress Notch signaling by reducing the expression of NOTCH 1 and jagged one.
Conclusions: Our findings indicate that the andrographolide constraints SW-480 cell growth through inhibition of the Notch signaling pathway.

Andrographolide Inhibits Proliferation of Colon Cancer SW-480 Cells via Downregulating Notch Signaling Pathway
Andrographolide Inhibits Proliferation of Colon Cancer SW-480 Cells via Downregulating Notch Signaling Pathway

MicroRNA-133b exacerbate atherosclerosis by activating the Notch signaling pathway1

The purpose of this study was to determine the effect of miR-133b in atherosclerosis (AS). A rat model of AS (AS Group) was established, and serum levels of total cholesterol, triglycerides, high-density lipoprotein cholesterol and low-density lipoprotein (LDL) cholesterol were detected. The thoracic aorta tissue was subjected to hematoxylin and eosin staining for pathological examination. Mice were intravenously injected with microRNA (mir) -133b mimic (group miR-133b mimic + US) and miR-133b mimic negative control (miR-133b group NC + US).

Normal mice given the name of Sham group. reconstruction parameters of vascular, inflammatory factors Ratio Collagen / Vascular Area (CA / CVA) and serum of mice in each group were detected. MRNA expression was measured by quantitative reverse transcription-PCR and protein expression was determined by western blot analysis. An in vitro model of the US induced in vascular smooth muscle cells (VSMC) using oxidized (ox) -LDL. CCK-8 and wound healing tests used to detect cell proliferation and migration.

Compared with Sham group, the rats of the US group, AS + miR-133b mimic NC group and the group AS + miR-133b have a higher intima thickness (IT), tumor necrosis factor (TNF) -α and monocytes kemoatraktan protein (MCP) -1 level, as well as increased expression of Notch1 and Jagged1; and they have a lower medial thickness (MT), the ratio CA / CVA and Notch3 expression (all P <0.05).

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In addition, miR-133b mimic promoted proliferation and migration, Notch1 and Jagged1 upregulated and downregulated Notch3 in VSMC ox-LDL-induced. Taken together, miR-133b worsen the US by activating Notch signaling pathway, which could serve as a potential target for the treatment of AS.

Anthony

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