Evaluation of the Anti-Tumoural and Immune Modulatory Activity of Natural Products by Flow Cytometry

Studies on the Identification of Constituents in Ethanol Extract of Radix Glycyrrhizae and Their Anti-Primary Hepatoma Cell Susceptibility

Journal of Chemistry ◽

10.1155/2014/134379 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5

Author(s):

Jie Liu

Keyword(s):

Flow Cytometry ◽

Natural Products ◽

Cell Cycle Progression ◽

Chemical Constituents ◽

Hepatoma Cell ◽

Ethanol Extract ◽

Hepatoma Cells ◽

Primary Hepatoma ◽

Isolation And Purification ◽

Radix Glycyrrhizae

The objective of this paper is to study the chemical constituents of Radix Glycyrrhizae and to apply the resulting natural products in the study of drug susceptibility of hepatoma cells so as to provide a scientific basis for quality standards and clinical application of medicinal Radix Glycyrrhizae. Chromatographic materials were used for isolation and purification; structural identification was performed based on physicochemical properties and spectral data. MTT colorimetry was used to detect the proliferation inhibition rate against primary hepatoma cells by natural products, and flow cytometry was used to detect the changes in cell cycle progression. Five compounds were isolated and identified, namely, liquiritigenin (1), liquiritin (2), isoliquiritigenin (3), betulinic acid (4), and oleanolic acid (5). In the study, 5-FU (5-fluorouracil) is used as a positive control to the hepatoma cells. Primary hepatoma cells were highly susceptible to 5-FU and liquiritigenin, both of which markedly inhibited the proliferation of hepatoma cells; flow cytometry results showed an increase in G0/G1 phase cells, a decrease in S phase cells, and a relative increase in G2/M phase cells. Primary hepatoma cells are highly susceptible to liquiritigenin, a natural product; the testing of tumor cell susceptibility is of important significance to the improvement of therapeutic effect of cancer.

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The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

10.26686/wgtn.17029535.v1 ◽

2021 ◽

Author(s):

◽

Lauren Taylor

Keyword(s):

Flow Cytometry ◽

Natural Products ◽

Morphological Analysis ◽

Accurate Information ◽

Biologically Relevant ◽

Multiple Components ◽

Selection Protocol ◽

Cellular Components ◽

Compound Concentration ◽

Product Compound

Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory. First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods. Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete. Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.

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Morphological Effects of Natural Products on Schizosaccharomyces pombe Measured by Imaging Flow Cytometry

Natural Products and Bioprospecting ◽

10.1007/s13659-014-0004-8 ◽

2014 ◽

Vol 4 (1) ◽

pp. 27-35 ◽

Cited By ~ 4

Author(s):

Joel Heisler ◽

Lindsay Elvir ◽

Farah Barnouti ◽

Erica Charles ◽

Tom D. Wolkow ◽

...

Keyword(s):

Flow Cytometry ◽

Natural Products ◽

Schizosaccharomyces Pombe ◽

Imaging Flow Cytometry

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The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

10.26686/wgtn.17029535 ◽

2021 ◽

Author(s):

◽

Lauren Taylor

Keyword(s):

Flow Cytometry ◽

Natural Products ◽

Morphological Analysis ◽

Accurate Information ◽

Biologically Relevant ◽

Multiple Components ◽

Selection Protocol ◽

Cellular Components ◽

Compound Concentration ◽

Product Compound

Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory. First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods. Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete. Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.

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6-Shogaol Suppresses the Progression of Liver Cancer via the Inactivation of Wnt/β-Catenin Signaling by Regulating TLR4

The American Journal of Chinese Medicine ◽

10.1142/s0192415x21500968 ◽

2021 ◽

Vol 49 (08) ◽

pp. 2033-2048

Author(s):

Yi Zhang ◽

Jingjing Wang ◽

Yong Qu ◽

Yunzhong Chen

Keyword(s):

Flow Cytometry ◽

Natural Products ◽

Liver Cancer ◽

Cancer Cells ◽

Western Blotting ◽

Mtt Assay ◽

Malignant Tumor ◽

Cell Apoptosis ◽

Inhibitory Effect ◽

Liver Cancer Cells

Liver cancer is a gastrointestinal malignant tumor with high lethality. The prognosis of liver cancer remains poor. Compounds derived from natural products have been confirmed to alleviate the progression of various diseases, including cancers. Additionally, 6-Shogaol has been reported to induce apoptosis in liver cancer cells. However, the mechanism by which 6-shogaol regulates apoptosis in liver cancer cells remains unclear. To investigate the function of 6-shogaol in liver cancer, RT–qPCR and western blotting were used to detect the expression of TLR4 and FOXO3a in liver cancer cells, respectively. The OD value of liver cancer cells was measured using the MTT assay. Flow cytometry was used to measure cell apoptosis. 6-Shogaol inhibited the growth of liver cancer cells. TLR4 and Wnt/[Formula: see text]-catenin were upregulated in liver cancer cells, and FOXO3a was inactivated, but 6-Shogaol reversed the expression of TLR4, Wnt/[Formula: see text]-catenin and FOXO3a in liver cancer cells. Additionally, TLR4 overexpression partially reversed the inhibitory effect of 6-shogaol on the progression of liver cancer cells via Wnt/[Formula: see text]-catenin signaling. Furthermore, the 6-shogaol-induced increase in FOXO3a expression in liver cancer was notably suppressed by TLR4 or Wnt/[Formula: see text]-catenin upregulation. Thus, 6-Shogaol suppresses the progression of liver cancer by mediating Wnt/[Formula: see text]-catenin signaling and is a potential agent for the treatment of liver cancer.

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