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Thomas Smith
Thomas Smith

BГ i 4 !FREE!

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BГ i 4

Momordica charantia, commonly known as bitter gourd (BG) or bitter melon, belongs to the family of Cucurbitaceae and is cultivated throughout the world, traditionally been used in folk medicines in cases of malaria, peptic ulcers, kidney stone, eczema14 to name a few and is commonly used in the treatment of type II diabetes15,16. In addition, extracts from different parts of BG possessed anticancer properties. MAP30, momordica anti-HIV protein, inhibited proliferation of MDA-MB-231 breast cancer cells and increased survival in MDA-MB-231 breast cancer mice xenografts17, methanolic extract of BG led to reduced proliferation and activated autophagy in CRC18 and sensitized CRC to the cytotoxic effect of doxorubicin19 while BG lectin has been shown to induce G2/M cell cycle arrest, autophagy and apoptosis in hepatocellular cancer20. These previous studies support the idea on the potential therapeutic use of BG and its biologically active constituents.

In the present study, we have devised a simple and effective procedure to isolate low-molecular weight protease inhibitor (BG-4) from BG and demonstrate its potential therapeutic property against colon cancer cells. The objectives were to characterize this novel anticancer peptide and to assess its capability to affect the growth of two human colon cancer cell lines, HCT-116 and HT-29. We first report here the discovery of a novel anticancer peptide from Momordica charantia by utilizing human colon cancer cells. We showed that BG-4 is very potent in inhibiting proliferation of both colon cancer cells, promoted apoptosis as measured by flow cytometry and microscopy study and the mechanism of action involved is through downregulation of antiapoptotic proteins Bcl-2 and XIAP, upregulation of proapoptotic proteins Bax and caspase-3 and modification of cell cycle proteins p21 and cyclin dependent kinase 2 (CDK2).

(a) Representative images from at least 3 independent experiments with 6 random fields per experiment taken from untreated and BG-4 treated HCT-116 human colon cancer cells. (b) Representative images from at least 3 independent experiments with 6 random fields per experiment taken from untreated and BG-4 treated HT-29 human colon cancer cells. Increased green fluorescence in BG-4 treated cells indicates that cells mitochondria became permeable to Annexin V, a characteristics of cells undergoing apoptosis.

We report for the first time the biological activity of a newly isolated and identified bioactive peptide from Momordica charantia we called BG-4. BG-4 was easily extracted using different concentrations of ethanol and can be purified from the rest of BG seed proteins by 70% ethanol as evidenced by electrophoresis profile of different BG seed aqueous ethanol extracts (Fig. 1a), a method that can be easily performed for the potential application of BG-4 as an anticancer agent. Amino acid sequencing of the purified peptide indicated that BG-4 amino acid sequences matched with the sequences of a protease inhibitor found in Momordica charantia (Table 1), interestingly all ethanolic protein extracts from Momordica charantia seed inhibited trypsin activity higher than purified soybean Kunitz trypsin inhibitor. The 70% ethanol extract containing purified BG-4 peptide inhibited trypsin activity 8.6 times higher than purified soybean trypsin inhibitor. Another interesting observation showed that increasing ethanol concentrations which is equivalent to increasing purity of BG-4 resulted in increased trypsin inhibitory activity (Fig. 1b) indicating that BG-4 is the responsible constituent for trypsin inhibition. Previous reports have shown that trypsin inhibitors from food sources possessed anticancer potential, the most studied are the Bowman-Birk inhibitor and Kunitz trypsin inhibitor from soybean22,23. Hence we tested BG-4 for its potential anticancer activities using two different human colon cancer cells.

In summary, we first report here a simple method for the isolation and purification of a novel anticancer peptide BG-4 from Momordica charantia. In addition, this study reports for the first time the capability of BG-4 to activate apoptosis in human colon cancer cells. Our findings support the idea for the potential use of BG-4 as a colon cancer therapeutic agent and must be studied using in vivo models of colon carcinogenesis.

How to cite this article: Dia, V. P. and Krishnan, H. B. BG-4, a novel anticancer peptide from bitter gourd (Momordica charantia), promotes apoptosis in human colon cancer cells. Sci. Rep. 6, 33532; doi: 10.1038/srep33532 (2016).

We thank Bo Song for performing the trypsin inhibitor assays. This work was supported by a start-up fund to VPD by Tennessee Agricultural Experiment Station. Product names are necessary to report factually on available data; however, USDA neither guarantees nor warrants the standard of product, and the use of the name by the USDA implies no approval of the product to the exclusion of others that may be suitable.

V.P.D. performed the experiments on the anti-colon cancer potential. H.B.K. performed the experiments on the isolation and characterization of BG-4 and trypsin inhibitory assay. V.P.D. and H.B.K. wrote the manuscript, contributed to the experimental design, interpretation of results and to the final editing of the manuscript.

**Society of Automotive Engineers (SAE) J1704It meets or exceeds Federal Motor Vehicle Safety Standard (FMVSS) No. 116 (DOT 4) and surpasses Society of Automotive Engineers (SAE) J1704 specifications.

Clean all dirt around the cover on the master cylinder before removing. Fill unit to level specified by manufacturer. Replace cover. Check fluid level once a month. To bleed the system, refer to the service manual. Do not reuse brake fluid from bleeding operation. CAUTION: Never wash cylinders or parts with gasoline, kerosene, or oil. Avoid spilling fluid on car finish or brake lining.

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Yes, our spray paint, paint pens and touch up bottles are ideal for touching up parts of all kinds, including powder coatings. However, keep in mind that our version of Munsell 7.5bg 7/4 is based on the color swatch contained in the official RAL color book. The color and sheen of your specific production parts may be different. If you need a touch up application, we can create a touch up solution precisely matching your specific parts.

Learn more about our color matching service. What are the LRV, HEX, and RGB values for the Munsell 7.5bg 7/4?The RGB values for Munsell 7.5bg 7/4 are 129, 186, 180 and the HEX code is #81BAB4. The LRV for Munsell 7.5bg 7/4 is 43.27. The LRV stands for Light Reflectance Value and measures the percentage of light that a color reflects. Learn more about using RGB and Hex codes for matching colors and Light Reflectance Values. What paint type is used to make spray paint matched to Munsell 7.5bg 7/4? And how many spray cans do I need?MyPerfectColor uses an acrylic enamel which is a fast-drying durable coating suitable for interior or exterior use. MyPerfectColor custom spray paint matched to Munsell 7.5bg 7/4 enables you to conveniently achieve a professional spray-smooth finish in any color in any sheen. It sticks well to most surfaces including metal, plastics, powder-coatings, cabinets and primed or previously painted wood.

The MyPerfectColor 11oz spray will cover about 20 square feet per coat. Keep in mind that it is difficult to gauge spray paint need as the coverage is highly dependent on how it is applied. Learn more about how spray paint works at MyPerfectColor.

Munsell Color continues the legacy of Albert H. Munsell and his dedication to helping people communicate color easily and accurately. Their aim is to provide you with effective color tools, tips, and techniques based on Munsell color theory.

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