Limits...
Combined ginger extract & Gelam honey modulate Ras/ERK and PI3K/AKT pathway genes in colon cancer HT29 cells.

Tahir AA, Sani NF, Murad NA, Makpol S, Ngah WZ, Yusof YA - Nutr J (2015)

Bottom Line: The cells were divided into 4 groups: the first group represents HT29 cells without treatment, the second and third groups were cells treated singly with either ginger or Gelam honey, respectively, and the last group represents cells treated with ginger and Gelam honey combined.Cell death in response to the combined ginger and Gelam honey treatment was associated with the stimulation of early apoptosis (upregulation of caspase 9 and IκB genes) accompanied by downregulation of the KRAS, ERK, AKT, Bcl-xL, NFkB (p65) genes in a synergistic manner.In conclusion, the combination of ginger and Gelam honey may be an effective chemopreventive and therapeutic strategy for inducing the death of colon cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia. analbaik@gmail.com.

ABSTRACT

Background: The interconnected Ras/ERK and PI3K/AKT pathways play a central role in colorectal tumorigenesis, and they are targets for elucidating mechanisms involved in attempts to induce colon cancer cell death. Both ginger (Zingiber officinale) and honey have been shown to exhibit anti-tumor and anti-inflammation properties against many types of cancer, including colorectal cancer. However, there are currently no reports showing the combined effect of these two dietary compounds in cancer growth inhibition. The aim of this study was to evaluate the synergistic effect of crude ginger extract and Gelam honey in combination as potential cancer chemopreventive agents against the colorectal cancer cell line HT29.

Methods: The cells were divided into 4 groups: the first group represents HT29 cells without treatment, the second and third groups were cells treated singly with either ginger or Gelam honey, respectively, and the last group represents cells treated with ginger and Gelam honey combined.

Results: The results of MTS assay showed that the IC50 of ginger and Gelam honey alone were 5.2 mg/ml and 80 mg/ml, respectively, whereas the IC50 of the combination treatment was 3 mg/ml of ginger plus 27 mg/ml of Gelam honey with a combination index of < 1, suggesting synergism. Cell death in response to the combined ginger and Gelam honey treatment was associated with the stimulation of early apoptosis (upregulation of caspase 9 and IκB genes) accompanied by downregulation of the KRAS, ERK, AKT, Bcl-xL, NFkB (p65) genes in a synergistic manner.

Conclusions: In conclusion, the combination of ginger and Gelam honey may be an effective chemopreventive and therapeutic strategy for inducing the death of colon cancer cells.

Show MeSH

Related in: MedlinePlus

Effects of single and combined treatments with ginger & Gelam honey on expression of Ras/ERK & PI3K/AKT pathway genes in HT29 cells. RNA was isolated, reverse transcribed to cDNA, and then amplified by a real-time PCR detection system to measure mRNA levels of GAPDH, KRAS(A), ERK(B)AKT(C), Bcl-xL(D), caspase 9(E), NFĸB/p65(F), IĸBα(G). Target genes were normalized to GAPDH. The data are presented as the mean ± SEM from three independent experiments. a(p < 0.05) significant compared to control, b(p < 0.05) significant compared to 2.5 mg/ml of ginger, c(p < 0.05) significant compared to 5.0 mg/ml of ginger, d(p < 0.05) significant compared to 7.5 mg/ml of ginger, e(p < 0.05) significant compared to 40 mg/ml of Gelam honey, f(p < 0.05) significant compared to 80 mg/ml of Gelam honey, g(p < 0.05) significant compared to 100 mg/ml of Gelam honey, h(p < 0.05) significant compared to combination of 3 mg/ml ginger and 10 mg/ml Gelam honey, i(p < 0.05) significant compared to combination of 3 mg/ml ginger and 30 mg/ml Gelam honey.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4390091&req=5

Fig4: Effects of single and combined treatments with ginger & Gelam honey on expression of Ras/ERK & PI3K/AKT pathway genes in HT29 cells. RNA was isolated, reverse transcribed to cDNA, and then amplified by a real-time PCR detection system to measure mRNA levels of GAPDH, KRAS(A), ERK(B)AKT(C), Bcl-xL(D), caspase 9(E), NFĸB/p65(F), IĸBα(G). Target genes were normalized to GAPDH. The data are presented as the mean ± SEM from three independent experiments. a(p < 0.05) significant compared to control, b(p < 0.05) significant compared to 2.5 mg/ml of ginger, c(p < 0.05) significant compared to 5.0 mg/ml of ginger, d(p < 0.05) significant compared to 7.5 mg/ml of ginger, e(p < 0.05) significant compared to 40 mg/ml of Gelam honey, f(p < 0.05) significant compared to 80 mg/ml of Gelam honey, g(p < 0.05) significant compared to 100 mg/ml of Gelam honey, h(p < 0.05) significant compared to combination of 3 mg/ml ginger and 10 mg/ml Gelam honey, i(p < 0.05) significant compared to combination of 3 mg/ml ginger and 30 mg/ml Gelam honey.

Mentions: Modulation of the gene expression of KRAS, ERK, AKT, Bcl-xL, caspase 9, p65 and IκBα involved in the Ras/ERK and PI3K/AKT pathways was investigated with ginger and Gelam honey. The gene expression levels of KRAS, ERK and AKT involved in the Ras/ERK/PI3K/AKT pathways were high in control cells (Figure 4A,B, and C). Treatment with ginger or Gelam honey alone resulted in the downregulation of the KRAS gene compared to control. However, a synergistic downregulation of KRAS was observed with combination treatment, especially at the dose of 3 mg/ml ginger in combination with either 30 or 50 mg/ml honey. As shown in Figure 4B, treatment with ginger alone downregulated expression of the ERK gene in a dose dependent manner, but combination of ginger with Gelam honey showed a greater effect on downregulating ERK gene expression compared to ginger or Gelam honey treatment alone. Treatment with either ginger or Gelam honey was found to downregulate AKT gene expression independently of the dose (Figure 4C). However, combination of both ginger and Gelam honey yielded higher downregulation of AKT gene expression at a much lower concentration compared to either treatment alone. The expression of Bcl-xL, an anti-apoptosis gene that was upregulated in the control group, was downregulated by either individual treatment, whereas combination treatment showed an even greater effect (Figure 4D). Conversely, expression of the pro-apoptosis gene caspase 9 (Figure 4E), which was downregulated in the untreated control group, was upregulated in treatments with high concentrations of ginger or Gelam honey alone, and a synergistic upregulation of caspase 9 was observed with the combined treatment at a much lower dose. As seen in Figure 4F, the inflammatory gene NFκB (p65), which is upregulated in cancer cells, was downregulated in treatments with ginger and Gelam honey alone or in combination, whereas the inhibitory gene IκBα (Figure 4G), which is downregulated in cancer cell lines, was highly upregulated with the combination treatment.Figure 4


Combined ginger extract & Gelam honey modulate Ras/ERK and PI3K/AKT pathway genes in colon cancer HT29 cells.

Tahir AA, Sani NF, Murad NA, Makpol S, Ngah WZ, Yusof YA - Nutr J (2015)

Effects of single and combined treatments with ginger & Gelam honey on expression of Ras/ERK & PI3K/AKT pathway genes in HT29 cells. RNA was isolated, reverse transcribed to cDNA, and then amplified by a real-time PCR detection system to measure mRNA levels of GAPDH, KRAS(A), ERK(B)AKT(C), Bcl-xL(D), caspase 9(E), NFĸB/p65(F), IĸBα(G). Target genes were normalized to GAPDH. The data are presented as the mean ± SEM from three independent experiments. a(p < 0.05) significant compared to control, b(p < 0.05) significant compared to 2.5 mg/ml of ginger, c(p < 0.05) significant compared to 5.0 mg/ml of ginger, d(p < 0.05) significant compared to 7.5 mg/ml of ginger, e(p < 0.05) significant compared to 40 mg/ml of Gelam honey, f(p < 0.05) significant compared to 80 mg/ml of Gelam honey, g(p < 0.05) significant compared to 100 mg/ml of Gelam honey, h(p < 0.05) significant compared to combination of 3 mg/ml ginger and 10 mg/ml Gelam honey, i(p < 0.05) significant compared to combination of 3 mg/ml ginger and 30 mg/ml Gelam honey.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4390091&req=5

Fig4: Effects of single and combined treatments with ginger & Gelam honey on expression of Ras/ERK & PI3K/AKT pathway genes in HT29 cells. RNA was isolated, reverse transcribed to cDNA, and then amplified by a real-time PCR detection system to measure mRNA levels of GAPDH, KRAS(A), ERK(B)AKT(C), Bcl-xL(D), caspase 9(E), NFĸB/p65(F), IĸBα(G). Target genes were normalized to GAPDH. The data are presented as the mean ± SEM from three independent experiments. a(p < 0.05) significant compared to control, b(p < 0.05) significant compared to 2.5 mg/ml of ginger, c(p < 0.05) significant compared to 5.0 mg/ml of ginger, d(p < 0.05) significant compared to 7.5 mg/ml of ginger, e(p < 0.05) significant compared to 40 mg/ml of Gelam honey, f(p < 0.05) significant compared to 80 mg/ml of Gelam honey, g(p < 0.05) significant compared to 100 mg/ml of Gelam honey, h(p < 0.05) significant compared to combination of 3 mg/ml ginger and 10 mg/ml Gelam honey, i(p < 0.05) significant compared to combination of 3 mg/ml ginger and 30 mg/ml Gelam honey.
Mentions: Modulation of the gene expression of KRAS, ERK, AKT, Bcl-xL, caspase 9, p65 and IκBα involved in the Ras/ERK and PI3K/AKT pathways was investigated with ginger and Gelam honey. The gene expression levels of KRAS, ERK and AKT involved in the Ras/ERK/PI3K/AKT pathways were high in control cells (Figure 4A,B, and C). Treatment with ginger or Gelam honey alone resulted in the downregulation of the KRAS gene compared to control. However, a synergistic downregulation of KRAS was observed with combination treatment, especially at the dose of 3 mg/ml ginger in combination with either 30 or 50 mg/ml honey. As shown in Figure 4B, treatment with ginger alone downregulated expression of the ERK gene in a dose dependent manner, but combination of ginger with Gelam honey showed a greater effect on downregulating ERK gene expression compared to ginger or Gelam honey treatment alone. Treatment with either ginger or Gelam honey was found to downregulate AKT gene expression independently of the dose (Figure 4C). However, combination of both ginger and Gelam honey yielded higher downregulation of AKT gene expression at a much lower concentration compared to either treatment alone. The expression of Bcl-xL, an anti-apoptosis gene that was upregulated in the control group, was downregulated by either individual treatment, whereas combination treatment showed an even greater effect (Figure 4D). Conversely, expression of the pro-apoptosis gene caspase 9 (Figure 4E), which was downregulated in the untreated control group, was upregulated in treatments with high concentrations of ginger or Gelam honey alone, and a synergistic upregulation of caspase 9 was observed with the combined treatment at a much lower dose. As seen in Figure 4F, the inflammatory gene NFκB (p65), which is upregulated in cancer cells, was downregulated in treatments with ginger and Gelam honey alone or in combination, whereas the inhibitory gene IκBα (Figure 4G), which is downregulated in cancer cell lines, was highly upregulated with the combination treatment.Figure 4

Bottom Line: The cells were divided into 4 groups: the first group represents HT29 cells without treatment, the second and third groups were cells treated singly with either ginger or Gelam honey, respectively, and the last group represents cells treated with ginger and Gelam honey combined.Cell death in response to the combined ginger and Gelam honey treatment was associated with the stimulation of early apoptosis (upregulation of caspase 9 and IκB genes) accompanied by downregulation of the KRAS, ERK, AKT, Bcl-xL, NFkB (p65) genes in a synergistic manner.In conclusion, the combination of ginger and Gelam honey may be an effective chemopreventive and therapeutic strategy for inducing the death of colon cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia. analbaik@gmail.com.

ABSTRACT

Background: The interconnected Ras/ERK and PI3K/AKT pathways play a central role in colorectal tumorigenesis, and they are targets for elucidating mechanisms involved in attempts to induce colon cancer cell death. Both ginger (Zingiber officinale) and honey have been shown to exhibit anti-tumor and anti-inflammation properties against many types of cancer, including colorectal cancer. However, there are currently no reports showing the combined effect of these two dietary compounds in cancer growth inhibition. The aim of this study was to evaluate the synergistic effect of crude ginger extract and Gelam honey in combination as potential cancer chemopreventive agents against the colorectal cancer cell line HT29.

Methods: The cells were divided into 4 groups: the first group represents HT29 cells without treatment, the second and third groups were cells treated singly with either ginger or Gelam honey, respectively, and the last group represents cells treated with ginger and Gelam honey combined.

Results: The results of MTS assay showed that the IC50 of ginger and Gelam honey alone were 5.2 mg/ml and 80 mg/ml, respectively, whereas the IC50 of the combination treatment was 3 mg/ml of ginger plus 27 mg/ml of Gelam honey with a combination index of < 1, suggesting synergism. Cell death in response to the combined ginger and Gelam honey treatment was associated with the stimulation of early apoptosis (upregulation of caspase 9 and IκB genes) accompanied by downregulation of the KRAS, ERK, AKT, Bcl-xL, NFkB (p65) genes in a synergistic manner.

Conclusions: In conclusion, the combination of ginger and Gelam honey may be an effective chemopreventive and therapeutic strategy for inducing the death of colon cancer cells.

Show MeSH
Related in: MedlinePlus