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Immunomodulatory and anticancer potential of Gan cao (Glycyrrhiza uralensis Fisch.) polysaccharides by CT-26 colon carcinoma cell growth inhibition and cytokine IL-7 upregulation in vitro.

Ayeka PA, Bian Y, Mwitari PG, Chu X, Zhang Y, Uzayisenga R, Otachi EO - BMC Complement Altern Med (2016)

Bottom Line: The ED50 was found to be 100 μg/ml.The effect of the polysaccharides were found to be molecular weight depended, with low molecular weight having a profound effect compared to high molecular weight and total crude extract.Therefore, immunomodulation is a possible mode of action of the polysaccharides in cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Tianjin University of Traditional Chinese Medicine, 312 Anshan Western Road, Nankai District, Tianjin, 300193, People's Republic of China.

ABSTRACT

Background: Chinese licorice, (Glycyrrhiza uralensis Fisch.) is one of the commonly prescribed herbs in Traditional Chinese Medicine (TCM). Gancao, as commonly known in China, is associated with immune-modulating and anti-tumor potential though the mechanism of action is not well known. In this study, we investigated the in vitro immunomodulatory and antitumor potential of Glycyrrhiza uralensis polysaccharides fractions of high molecular weight (fraction A), low molecular weight (fraction B) and crude extract (fraction C).

Methods: Cell proliferation and cytotoxicity was investigated using Cell Counting kit 8 (CCK-8) on Intestinal epithelial cell line (IEC-6) and Colon carcinoma cell line (CT-26). IL-7 gene expression relative to GAPDH was analysed using Real time PCR. The stimulation and viability of T lymphocytes was determined by Trypan blue exclusion assay.

Results: G.uralensis polysaccharides did not inhibit proliferation of IEC-6 cells even at high concentration. The ED50 was found to be 100 μg/ml. On the other hand, the polysaccharides inhibited the proliferation of cancer cells (CT-26) at a concentration of ≤50 μg/ml. Within 72 h of treatment with the polysaccharides, expression of IL-7 gene was up-regulated over 2 times. It was also noted that, IEC-6 cells secrete IL-7 cytokine into media when treated with G.uralensis polysaccharides. The secreted IL-7 stimulated proliferation of freshly isolated T lymphocytes within 6 h. The effect of the polysaccharides were found to be molecular weight depended, with low molecular weight having a profound effect compared to high molecular weight and total crude extract.

Conclusion: Our findings indicate that G.uralensis polysaccharides especially those of low molecular weight have a potential as anticancer agents. Of great importance, is the ability of the polysaccharides to up-regulate anticancer cytokine IL-7, which is important in proliferation and maturation of immune cells and it is associated with better prognosis in cancer. Therefore, immunomodulation is a possible mode of action of the polysaccharides in cancer therapy.

No MeSH data available.


Related in: MedlinePlus

Expression of IL 7mRNA at (a) 3 h and (b) 72 h. 100 μg/ml of each polysaccharide fraction was added to IEC-6 cells and incubated for 3 and 72 h respectively. The normalized relative expression of IL-7 mRNA was analyzed by IQ5 Real-time PCR software. Data presented as mean ± S.E.M. *P < 0.05 versus untreated (control) of three experiments
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Fig3: Expression of IL 7mRNA at (a) 3 h and (b) 72 h. 100 μg/ml of each polysaccharide fraction was added to IEC-6 cells and incubated for 3 and 72 h respectively. The normalized relative expression of IL-7 mRNA was analyzed by IQ5 Real-time PCR software. Data presented as mean ± S.E.M. *P < 0.05 versus untreated (control) of three experiments

Mentions: To investigate the immunomodulatory effect of G.uralensis polysaccharides through upregulation of IL-7 gene, the expression level of IL-7 in licorice polysaccharide treated IEC-6 cells was determined. The IEC-6 cells treated with G.uralensis polysaccharides showed up-regulation of the IL-7 gene after 3 h and the upregulation remained elevated upto 72 h of treated. The up regulation of these cytokine was depended on the molecular weight of the extracts, going up by over two times with fraction B (Fig. 3). The up-regulation of the cytokine was not time dependent, at 3 and 72 h, the up-regulation was not significantly different (Fig. 3a and b).Fig. 3


Immunomodulatory and anticancer potential of Gan cao (Glycyrrhiza uralensis Fisch.) polysaccharides by CT-26 colon carcinoma cell growth inhibition and cytokine IL-7 upregulation in vitro.

Ayeka PA, Bian Y, Mwitari PG, Chu X, Zhang Y, Uzayisenga R, Otachi EO - BMC Complement Altern Med (2016)

Expression of IL 7mRNA at (a) 3 h and (b) 72 h. 100 μg/ml of each polysaccharide fraction was added to IEC-6 cells and incubated for 3 and 72 h respectively. The normalized relative expression of IL-7 mRNA was analyzed by IQ5 Real-time PCR software. Data presented as mean ± S.E.M. *P < 0.05 versus untreated (control) of three experiments
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4940688&req=5

Fig3: Expression of IL 7mRNA at (a) 3 h and (b) 72 h. 100 μg/ml of each polysaccharide fraction was added to IEC-6 cells and incubated for 3 and 72 h respectively. The normalized relative expression of IL-7 mRNA was analyzed by IQ5 Real-time PCR software. Data presented as mean ± S.E.M. *P < 0.05 versus untreated (control) of three experiments
Mentions: To investigate the immunomodulatory effect of G.uralensis polysaccharides through upregulation of IL-7 gene, the expression level of IL-7 in licorice polysaccharide treated IEC-6 cells was determined. The IEC-6 cells treated with G.uralensis polysaccharides showed up-regulation of the IL-7 gene after 3 h and the upregulation remained elevated upto 72 h of treated. The up regulation of these cytokine was depended on the molecular weight of the extracts, going up by over two times with fraction B (Fig. 3). The up-regulation of the cytokine was not time dependent, at 3 and 72 h, the up-regulation was not significantly different (Fig. 3a and b).Fig. 3

Bottom Line: The ED50 was found to be 100 μg/ml.The effect of the polysaccharides were found to be molecular weight depended, with low molecular weight having a profound effect compared to high molecular weight and total crude extract.Therefore, immunomodulation is a possible mode of action of the polysaccharides in cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Tianjin University of Traditional Chinese Medicine, 312 Anshan Western Road, Nankai District, Tianjin, 300193, People's Republic of China.

ABSTRACT

Background: Chinese licorice, (Glycyrrhiza uralensis Fisch.) is one of the commonly prescribed herbs in Traditional Chinese Medicine (TCM). Gancao, as commonly known in China, is associated with immune-modulating and anti-tumor potential though the mechanism of action is not well known. In this study, we investigated the in vitro immunomodulatory and antitumor potential of Glycyrrhiza uralensis polysaccharides fractions of high molecular weight (fraction A), low molecular weight (fraction B) and crude extract (fraction C).

Methods: Cell proliferation and cytotoxicity was investigated using Cell Counting kit 8 (CCK-8) on Intestinal epithelial cell line (IEC-6) and Colon carcinoma cell line (CT-26). IL-7 gene expression relative to GAPDH was analysed using Real time PCR. The stimulation and viability of T lymphocytes was determined by Trypan blue exclusion assay.

Results: G.uralensis polysaccharides did not inhibit proliferation of IEC-6 cells even at high concentration. The ED50 was found to be 100 μg/ml. On the other hand, the polysaccharides inhibited the proliferation of cancer cells (CT-26) at a concentration of ≤50 μg/ml. Within 72 h of treatment with the polysaccharides, expression of IL-7 gene was up-regulated over 2 times. It was also noted that, IEC-6 cells secrete IL-7 cytokine into media when treated with G.uralensis polysaccharides. The secreted IL-7 stimulated proliferation of freshly isolated T lymphocytes within 6 h. The effect of the polysaccharides were found to be molecular weight depended, with low molecular weight having a profound effect compared to high molecular weight and total crude extract.

Conclusion: Our findings indicate that G.uralensis polysaccharides especially those of low molecular weight have a potential as anticancer agents. Of great importance, is the ability of the polysaccharides to up-regulate anticancer cytokine IL-7, which is important in proliferation and maturation of immune cells and it is associated with better prognosis in cancer. Therefore, immunomodulation is a possible mode of action of the polysaccharides in cancer therapy.

No MeSH data available.


Related in: MedlinePlus