Antiproliferative, antimigratory, and anticlonogenic effects of Hedyotis diffusa, Panax ginseng, and their combination on colorectal cancer cell lines

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1 Journal of Herbs, Spices & Medicinal Plants ISSN: (Print) (Online) Journal homepage: Antiproliferative, antimigratory, and anticlonogenic effects of Hedyotis diffusa, Panax ginseng, and their combination on colorectal cancer cell lines KetLi Ho, Ai Peng Lew, Yee Ching Ong, Yun Qian Lew & Zhi Hang Wong To cite this article: KetLi Ho, Ai Peng Lew, Yee Ching Ong, Yun Qian Lew & Zhi Hang Wong (2018): Antiproliferative, antimigratory, and anticlonogenic effects of Hedyotis diffusa, Panax ginseng, and their combination on colorectal cancer cell lines, Journal of Herbs, Spices & Medicinal Plants, DOI: / To link to this article: Published online: 30 Jan Submit your article to this journal Article views: 8 View related articles View Crossmark data Full Terms & Conditions of access and use can be found at

2 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS Antiproliferative, antimigratory, and anticlonogenic effects of Hedyotis diffusa, Panax ginseng, and their combination on colorectal cancer cell lines KetLi Ho, Ai Peng Lew, Yee Ching Ong, Yun Qian Lew, and Zhi Hang Wong Research Laboratory, Department of Life Science, International Medical University, Bukit Jalil, Kuala Lumpur, Malaysia ABSTRACT Combinations of herbs are often used in traditional Chinese medicine (TCM) as the complex constituents of herbs could synergize in producing greater therapeutic efficacy. The combination of Hedyotis diffusa and Panax ginseng is commonly used to treat colorectal cancer in TCM. This study compared the antiproliferative, antimigratory, and anticlonogenic effects of these herbs when used individually and in combination. Results suggested that the efficacy of the combined herbs was not superior to that of the individual herbs in inhibiting cancer cell proliferation and migration. However, the combinations acted synergistically in inhibiting colony formation. ARTICLE HISTORY Received 28 June 2017 KEYWORDS Cell proliferation; cell migration; colony formation; combination index Introduction Colorectal cancer (CRC) is the second and third most commonly diagnosed cancer in female and male worldwide, respectively (12). Chemotherapy, radiotherapy, and surgery, the main conventional treatments for CRC (22,27) often caused adverse effects and impair patients quality of life (22,27). Therefore, many patients opt for complementary medicine as it is perceived as having fewer side effects compared to conventional medicine (11). The use of herbal decoction in traditional Chinese medicine (TCM) is based on the assumption that multiple constituents of herbs act synergistically in regulating the disharmony in the body. According to TCM theory, heat-induced toxin and stagnation of vital energy are responsible for colon carcinogenesis (5,6). Hence, Hedyotis diffusa and Panax ginseng which have heat clearing and vital energy restoration properties, respectively, are used to treat CRC (5,10,19,21,27). When used in combination, P. ginseng is believed to be able to reinforce the anticancer effects of H. diffusa in CRC patients (5,10). CONTACT KetLi Ho Research Laboratory, International Medical University, No. 126, Jalan Jalil Perkasa 19, Bukit Jalil, Kuala Lumpur, Malaysia. Color versions of one or more of the figures in the article can be found online at Taylor & Francis Group, LLC

3 2 K. HO ET AL. The anticancer effects of H. diffusa and P. ginseng have been previously reported. H. diffusa induced cell death (18,20), inhibited colony formation (20), cell migration (8,17), and arrested the cell cycle of colon carcinoma cells in G 0 /G 1 checkpoint with a concomitant reduction of cell population in S phase (26). Similarly, P. ginseng inhibited cell proliferation (9,25), inactivated antiapoptosis genes, induced cancer cell apoptosis (16), inhibited cell migration (3,13), and arrested the cell cycle (2). This study investigated the antiproliferative, antimigratory, and anticlonogenic effects of combination of H. diffusa and P. ginseng on colorectal cancer cell lines. Materials and methods Cell culture Human colorectal cancer cells HCC2998 and KM12 were obtained from the International Medical University Cell Bank and maintained in RPMI-1640 medium, supplemented with 10% (v/v) FBS and 0.5% (v/v) penicillin-streptomycin at 37 C in a humidified atmosphere containing 5% CO 2. H. diffusa and P. ginseng extracts The crude extracts of H. diffusa and P. ginseng were purchased from China Resource Sanjiu Medical & Pharmaceutical Company, separately dissolved in dimethyl sulfoxide (DMSO) to prepare a 400 mg ml 1 stock solution and stored at 4 C. The stock solution was diluted with RPMI-1640 medium to obtain the required concentrations. Antiproliferative effects of Hedyotis diffusa and Panax ginseng 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay was used to assess the antiproliferative effect of H. diffusa and P. ginseng. Briefly, cells/well were seeded in a 96-well plate and incubated overnight and then the cells were treated for 24 and 48 h. The final concentration of DMSO in the medium was <1%. At the end of the treatment, 20 µl of 5 mg ml 1 MTT solution was added into each well and the cells were incubated at 37 C for 4 h. After that, 100 μl of DMSO was added to dissolve the purple formazan crystals. The absorbance at 570 nm and 640 nm (reference wavelength) was measured by using a microplate reader (Tecan, Switzerland). The cell viability was calculated using the following equation: Mean absorbance of treated wells Cell viability ð% Þ ¼ Mean absorbance of control wells 100%

4 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS 3 The concentrations of herbs required to inhibit 50% of the cell viability (IC 50 ) were determined using nonlinear regression graphs. Antimigratory effects Wound healing assay was carried out to study the antimigratory effects of the herbs. Cells were seeded in a 24-well plates ( cells/well) and incubated until a confluent monolayer is formed. A sterilized ruler and a yellow pipette tip were used to scratch two straight lines on the confluent monolayer to create a cross. Then, the wells were washed twice with culture medium to remove the detached cells from the wells. Cells were treated with RPMI-1640 medium in eight different concentrations of extracts. The cells were incubated and visualized by using Nikon Eclipse inverted microscope Ti-U with 4 magnification and PhL filter (Nikon, Melville, NY). Images were captured using Nikon NIS-elements imaging software (Nikon, Melville, NY) immediately after the wounds were created (0 h) and at 24 h after the wounds were created. Migration of the cells was assessed through the recovery of the scratch. The closure of the wound was assessed by measuring the area between the wound edges, which was then calculated and quantified using Nikon NIS-elements imaging software (Nikon, Melville, NY). The percentage of wound closure was calculated with the following equation: Percentage of wound closure ð% Þ ¼ Area of the wound at 0 hour Area of the wound at 24 hours Area of the wound at 0 hour 100 Anticlonogenic effect of H. diffusa and P. ginseng Cells were seeded in 6-well plates at a density of 5000 cells/well with 3 ml of cultured media and incubated overnight. Subsequently, the cells were treated with different concentrations of herb extracts. After 8 days of incubation period, colonies were fixed with fixing solution (1:3 acetic acid: methanol) for 30 min and stained with 0.5% crystal violet for 2 h at room temperature. The number of colonies with more than 50 cells was counted manually under microscope. Additionally, the area of colonies was measured using Nikon NIS-Elements imaging software (Nikon, Melville, NY). Percentage of colony number and colony area were calculated as: Percentage of colony number ð% Þ : Number of colonies formed in treatment group Number of colonies formed in control group 100%

5 4 K. HO ET AL. Area of colonies formed in treatment group Percentage of colony area ð% Þ: Area of colonies formed in control group 100% Antiproliferative, antimigratory, and anticlonogenic effects of H. diffusa and P. ginseng combination The combinatory effect of H. diffusa and P. ginseng was determined (4) using a constant ratio design (Table 1). MTT assay, wound healing assay, and clonogenic assay were carried out as described earlier. Interaction between the herbs was examined with combination index (CI) analysis and the CI values were calculated using CompuSyn software (Biosoft, Cambridge, UK). Statistical analysis All experiments were carried out at least three times. One-way analysis of variance (ANOVA)followedbyTukeytest(p < 0.05) by using GraphPad Prism 7. Results Antiproliferative effects H. diffusa and P. ginseng used individually decreased cell proliferation in a time- and dose-dependent manner (Figure 1). The IC 50 obtained from HCC2998 cells after 24 and 48 h of treatment were (in µg ml 1 ) 2800 and 1000 respectively for H. diffusa and 1600 and 800 respectively for P. ginseng (Table 2). The antiproliferative effects of both H. diffusa and P. ginseng on KM12 were weaker compared with HCC2998 when applied individually. The IC 50 for H. diffusa was unattainable while the IC 50 for P. ginseng after 24 and 48 h Table 1. A Constant Ratio Experimental Design for Herb Combination Analysis. Hedyotis diffusa x (IC 50 ) x (IC 50 ) x (IC 50 ) x (IC 50 ) x (IC 50 ) 1 Panax 0 Control (f a ) 0 (f a ) 1 (f a ) 1 (f a ) 1 (f a ) 1 (f a ) 1 ginseng 0.25 x (IC 50 ) 2 (f a ) 2 (f a ) 1, x (IC 50 ) 2 (f a ) 2 (f a ) 1,2 1.0 x (IC 50 ) 2 (f a ) 2 (f a ) 1,2 2.0 x (IC 50 ) 2 (f a ) 2 (f a ) 1,2 4.0 x (IC 50 ) 2 (f a ) 2 (f a ) 1,2 (IC 50 ) 1 : Effective dose of Hedyotis diffusa which inhibited colorectal cancer cells proliferation, migration, and colony formation to 50% (IC 50 ) 2 : Effective dose of Panax ginseng which inhibited colorectal cancer cells proliferation, migration, and colony formation to 50% f a : Fraction affected by dose

6 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS 5 Figure 1. Antiproliferative effects of (A) Hedyotis diffusa on HCC2998, (B) Panax ginseng on HCC2998, (C) H. diffusa on KM12 and (D) P. ginseng on KM12. The data were normalized with negative control (treated with 1.0 % of DMSO as vehicle control). Each data point represents the mean ± standard deviation of triplicates. *p < 0.05; **p < 0.01; ***p < Table 2. IC 50 for Hedyotis diffusa and Panax ginseng Antiproliferative (AP) Antimigratory (AM) and Anticlonogenic (AC) Effects After the Colorectal Cancer Cells Were Treated for 24 h and 48 h. IC 50 (μg ml 1 ) HCC2998 KM12 24 h 48 h 24 h 48 h AP AM AC AP AM AC AP AM AC AP AM AC Hedyotis diffusa Panax ginseng were (in µg ml 1 ) 2000 and 1300 respectively (Table 2). The IC 50 for both H. diffusa and P. ginseng was required to study the antiproliferative effect of the herbs combination. As the IC 50 of H. diffusa on KM12 was unattainable, the combinatory effect in KM12 was not investigated. The herbs combination study carried out in HCC2998 showed that the herbs combination had a stronger antiproliferative effect than that of H. diffusa alone, but not different from the antiproliferative effect of P. ginseng alone (Figure 2). CI was employed to determine the nature of the interaction between the herbs. A CI value of <1, = 1, and >1 represented synergistic, additive, and antagonistic effect, respectively. The combination of H. diffusa and P. ginseng demonstrated an antagonistic effect (CI > 1) when F a (fraction of cells with low viability) increased (Figure 3).

7 6 K. HO ET AL. Figure 2. Dose effect curves of individual and combination of Hedyotis diffusa (HD) and Panax ginseng (PG) on HCC2998 cells. Fraction affected represents the fraction of cell with reduced viability. The dose effect curves of HD alone and combination with PG overlapped each other, suggesting that both have similar antiproliferative effects. Antimigratory effect of H. diffusa, P. ginseng, and their combination Wound healing assay was used to assess the antimigratory effect of H. diffusa and P. ginseng. H. diffusa decreased the migration of HCC2998 dose-dependently with the IC 50 of 700 µg ml 1 (Figure 4a and Table 2). However, P. ginseng increased HCC2998 cells migration dose-dependently instead of inhibiting it (Figure 4b). Thus, IC 50 value of P. ginseng on HCC2998 cell migration could Figure 3. Combination index (CI) vs. fraction affected (F a ) plot of the combination of Hedyotis diffusa and Panax ginseng on HCC2998. Fraction affected represents the fraction of cell with reduced viability. CI < 1, = 1 and > 1 indicates synergism, additive, and antagonism effect respectively.

8 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS 7 Figure 4. Antimigratory effects of (A) Hedyotis diffusa on HCC2998, (B) Panax ginseng on HCC2998, (C) H. diffusa on KM12 and (D) P. ginseng on KM12. The data were normalized with negative control (treated with 1.0 % of DMSO as vehicle control). Each data point represents the mean ± standard deviation of triplicates. *p < 0.05 compared to control. not be determined. Therefore, the combinatory effect of H. diffusa and P. ginseng on the migration of HCC2998 was not investigated. On the other hand, both H. diffusa and P. ginseng that were given individually resulted in a dose-dependent decrease in cell migration on KM12 cell line (Figure 4c and 4d). IC 50 for H. diffusa and P. ginseng was (in µg ml 1 ) 920 and 160 respectively (Table 2). For the antimigratory effect of the herbs combination, the dose effect curves (Figure 5) showed that P. ginseng used alone had higher effect than H. diffusa and the herbs combination. The combination of H. diffusa and P. ginseng demonstrated antagonistic effect except when the fraction affected (fraction of cells with reduced migration activity) was very high (Figure 6). Anticlonogenic effects H. diffusa and P. ginseng promoted dose-dependent reduction in colony formation capacity on both KM12 and HCC2998 CRC cell lines. As the concentration increased, both herb extracts decreased the number and area of colonies (Figures 7 10, Table 2). A dose respond effect was seen for H. diffusa and P. ginseng, whenused individually and in combination, on the colony formation of KM12 and

9 8 K. HO ET AL. Figure 5. Dose effect curves of individual and combination of Hedyotis diffusa (HD) and Panax ginseng (PG) on KM12 cells. Fraction affected represents the fraction of cell migration compared to negative control (treated with 1.0% of DMSO as vehicle control). Figure 6. Combination index (CI) vs fraction affected (F a ) plot of the combination of Hedyotis diffusa and Panax ginseng on KM12. Fraction affected (F a ) represents the fraction of cells with reduced migration activity. Dotted line in the graph CI = 1 indicates addictive effect. CI < 1, = 1 and > 1 indicates synergism, additive, and antagonism effects respectively. HCC2998 (Figures 11 and 12). Generally, the herbs combination showed better anticlonogenic effect than individual herbs. The combination of H. diffusa and P. ginseng demonstrated synergistic effect on both cell lines in all fraction affected (fraction of the cells with reduced clonogenic property) (Figures 13 and 14). Discussion Unregulated proliferation, migration, and colony formation are the universal hallmarks of cancerous cells (1,7,24). In this study, the

10 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS 9 Figure 7. Anticlonogenic effect of Hedyotis diffusa on HCC2998. Cells in control well were treated with 1.0% DMSO. Colonies with more than 50 cells were counted manually under the microscope. (A) Relative number of colonies compared to control. (B) Relative area of colonies compared to control. (C) Digital image showing colonies produced in HCC2998 following plating of 5000 cells and 8 d incubation. Each data point represents the mean ± standard deviation of triplicates. *p < 0.01 compared to control. Figure 8. Anticlonogenic effect of Hedyotis diffusa on KM12. Cells in control well were treated with 1.0% DMSO. Colonies with more than 50 cells were counted manually under the microscope. (A) Relative number of colonies compared to control (B) Relative area of colonies compared to control. (C) Digital image showing colonies produced in KM12 following plating of 5000 cells and 8 d incubation. Each data point represents the mean ± standard deviation of triplicates. *p < 0.01 compared to control. Figure 9. Anticlonogenic effect of Panax ginseng on HCC2998. Cells in control well were treated with 1.0% DMSO. Colonies with more than 50 cells were counted manually under the microscope. (A) Relative number of colonies compared to control. (B) Relative area of colonies compared to control. (C) Digital image showing colonies produced HCC2998 following plating of 5000 cells and 8 d incubation. Each data point represents the mean ± standard deviation of triplicates. *p < 0.01 compared to control. antiproliferative, antimigratory, and anticlonogenic effects of H. diffusa, P. ginseng, and their combination on two human colorectal cancer cell lines were examined.

11 10 K. HO ET AL. Figure 10. Anticlonogenic effect of Panax ginseng on KM12. Cells in control well were treated with 1.0% DMSO. Colonies with more than 50 cells were counted manually under the microscope. (A) Relative number of colonies compared to control. (B) Relative area of colonies compared to control. (C) Digital image showing colonies produced KM12 following plating of 5000 cells and 8 days incubation. Each data point represents the mean ± standard deviation of triplicates. * p < 0.01 compared to control. Figure 11. Dose effect curves of individual and combination of Hedyotis diffusa (HD) and PG, Panax ginseng (PG) on HCC2998. Fraction affected represents the fraction of colony formation compared to negative control (treated with 1.0% of DMSO as vehicle control). Previous studies on P. ginseng confirmed its antiproliferative effects on cancer cell lines in animal models (2,9,15,16,25); results are consistent with the findings of this study where P. ginseng demonstrated antiproliferative effect in a time- and dose-dependent manner on both cell lines (Figure 1b and 1d). However, despite the reported antiproliferative effect of H. diffusa in some previous studies (18,20), this study found that the antiproliferative effect of H. diffusa was cell line-specific. In this study, H. diffusa showed antiproliferative effect in a dose-dependent manner on HCC2998 (Figure 1a) but not on KM12 (Figure 1c), thus suggesting that the genetic make-up of the cells plays a crucial role in the response cancerous cells to herbal treatment. When the herbs were used in combination, synergistic effect was only shown

12 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS 11 Figure 12. Dose effect curves of individual and combination of herbs on KM12. Fraction affected represents the fraction of colony formation compared to negative control (treated with 1.0% of DMSO as vehicle control). HD, Hedyotis diffusa; PG, Panax ginseng; HD + PG, combination of Hedyotis diffusa and Panax ginseng. Figure 13. Combination index (CI) versus fraction affected (F a ) plot of the combination of Hedyotis diffusa and Panax ginseng on HCC2998. Fraction affected (F a ) represents the fraction of the cells with reduced clonogenic property compared to negative control (treated with 1.0% of DMSO as vehicle control). Dotted line in the graph CI = 1 indicates addictive effect. CI < 1, = 1 and > 1 indicates synergism, additive, and antagonism effect respectively. when the antiproliferative activity was low, while the antagonistic effect was portrayed when the antiproliferative activity increased (Figure 3). It should be noted that the combinatory effect at the high F a region (high antiproliferative effect) was more important than the low F a region (low antiproliferative effect) because incomplete eradication of colorectal cancer cells may lead to therapy resistance and cancer recurrence (4).

13 12 K. HO ET AL. Figure 14. Combination index (CI) versus fraction affected (F a ) plot of the combination of Hedyotis diffusa and Panax ginseng on KM12. Fraction affected (F a ) represents the fraction of cells with reduced clonogenic property compared to negative control (treated with 1.0% of DMSO as vehicle control). Dotted line in the graph CI = 1 indicates addictive effect. CI < 1, = 1 and > 1 indicates synergism, additive, and antagonism effect respectively. For antimigratory effect, both ginsenoside Rg3 (an active component of P. ginseng) (3,13,24) and H. diffusa extract (8,17,20) have been reported. In this study, P. ginseng showed more potent antimigratory effect on KM12 than H. diffusa (Figure 4). However, on HCC2998, P. ginseng did not show any antimigratory effect while H. diffusa showed a mild antimigratory effect (Figure 4). As the antimigratory effect of herbs combination can only be investigated when both herbs are effective, it was not carried out for HCC2998 in this study. When the herbs were used in combination on KM12, the herbs combination was found to show weaker antimigratory effect compared to P. ginseng alone (Figure 5). This was further supported by the CI plot (Figure 6) which showed that the antagonistic effect was seen when the herbs were exerting low to high antimigratory activities. Although synergistic effect was shown when the F a was very close to 1, it should be noted that complete inhibition of cell migration was unlikely to be achieved in vivo. Colony formation is a hallmark of carcinogenesis. Cancerous cells which retain their clonogenic ability could spread to different organs and thus result in metastasis (7). Both P. ginseng (14) and H. diffusa (20) have been reported to show strong anticlonogenic effects on cancer cell lines. These findings are consistent with the results of this study. Both H. diffusa and P. ginseng reduced the clonogenic ability of both cell lines in a dose-dependent manner (Figures 7 10). The IC 50 of P. ginseng was about twofold lower than that of

14 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS 13 H. diffusa (Table 2), indicating that P. ginseng had more potent anticlonogenic property than H. diffusa. When the herbs were used in combination, strong synergistic effects were observed on both cell lines (Figures 11 13). Due to the strong synergism, a complete inhibition of colony formation was achievable even at low concentrations of herb extracts. Acknowledgments This work was supported by International Medical University research grant (BP-I-01/13(31) 2016). References 1. Adams, J. M., and S. Cory The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene. 26(9): Cheng, H, S. Li, Y. Fan, X. Gao, M. Hao, J. Wang, X. Zhang, G. Tai, Y. Zhou Comparative studies of the antiproliferative effects of ginseng polysaccharides on HT- 29 human colon cancer cells. Med. Oncol. 28(1): Choo, M., H. Sakurai, D. Kim, and I. Saiki A ginseng saponin metabolite suppresses tumor necrosis factor-alpha-promoted metastasis by suppressing nuclear factor-kappa B signaling in murine colon cancer cells. Oncol. Rep. 19(3): Chou, T. C Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination Studies. Pharm. Rev. 58(3): Deng, S., B. Hu, and H. An Traditional Chinese medicinal syndromes and treatment in colorectal cancer. J. Cancer Ther. 3(06): Dorsher, P., and Z. Peng Chinese medicinal herbs use in managing cancer. In William C.S.Cho (Ed.), Supportive Cancer Care with Chinese Medicine. Springer. pp Gustin, D. M., and D. E. Brenner Chemoprevention of colon cancer: current status and future prospects. Cancer Metastasis Rev. 21(3 4): Han, Y., X. Chen, H. Gao, J. Ye, and C. Wang Physcion inhibits the metastatic potential of human colorectal cancer SW620 cells in vitro by suppressing the transcription factor SOX2. Acta Pharmacologica Sinica. 37(2): He, Bai-Cheng, J. Gao, X. Luo, J. Luo, J. Shen, L. Wang, Q. Zhou, Y. Wang, H. Luu, R. Haydon Ginsenoside Rg3 inhibits colorectal tumor growth through the downregulation of Wnt/β-catenin signaling. Int. J. Oncol. 38(2): Huang, R. H., F. Chen, G.Q. Lao, Z.Y. Shi, J.H. Luo, D.R. Liang, J.J. Chen Effect of Jian-Pi-Jie-Du decoction on the life quality of colorectal cancer patients in advanced stage. Hebei J. Trad. Chin. Med. 33(4): Hung, A., N. Kang, A. Bollom, J. L. Wolf, and A. Lembo Complementary and alternative medicine use is prevalent among patients with gastrointestinal diseases. Dig. Dis. Sci. 60(7): Jemal, A., F. Bray, M. M. Center, J. Ferlay, E. Ward, and D. Forman Global cancer statistics. CA: Cancer J. Clin. 61(2): Junmin, S., L. Hongxiang, L. Zhen, Y. Chao, and W. Chaojie Ginsenoside Rg3 inhibits colon cancer cell migration by suppressing nuclear factor Kappa B activity. J. Trad. Chin. Med. 35(4):

15 14 K. HO ET AL. 14. Kang, J. H., K. H. Song, J. K. Woo, M. H. Park, M. H. Rhee, C. Choi, and S. H. Oh Ginsenoside Rp1 from Panax ginseng exhibits anti-cancer activity by downregulation of the IGF-1R/Akt pathway in breast cancer cells. Plant Foods Human Nutr. 66(3): Kim, H., H. Lee, D. J. Kim, T. M. Kim, H. Moon, and H. Choi Panax ginseng exerts antiproliferative effects on rat hepatocarcinogenesis. Nutr. Res. 33(9): Kim, S. M., S. Y. Lee, D. Y. Yuk, D. C. Moon, S. S. Choi, and Y. Kim Inhibition of NF-κB by ginsenoside Rg3 enhances the susceptibility of colon cancer cells to docetaxel. Arch. Pharm. Res. 32(5): Kwak, H. J., M. J. Park, C. M. Park, S. Moon, D. H. Yoo, H.C. Lee, S.H. Lee, M.S. Kim, H.W. Lee, W.S. Shin, et al Emodin inhibits vascular endothelial growth factor-ainduced angiogenesis by blocking receptor-2 (KDR/Flk-1) phosphorylation. Int. J. Cancer. 118(11): Lee, J., K. Kwon, C. Cho, S. S. R. Han, and H. Yoo Advanced cancer cases treated with cultivated wild ginseng phamacopuncture. J. Acupuncture Meridian Stud. 3(2): Li, Q., X. Wang, A. Shen, Y. Zhang, Y. Chen, T. J. Sferra, J. Lin, and J. Peng Hedyotis diffusa willd overcomes 5-fluorouracil resistance in human colorectal cancer HCT-8/5-FU cells by downregulating the expression of P-glycoprotein and ATP-binding casette subfamily G member 2. Exp. Therap. Med. 10(5): Lin, J., Q. Li, H. Chen, H. Lin, Z. Lai, and J. Peng Hedyotis diffusa willd. Extract suppresses proliferation and induces apoptosis via IL-6-inducible STAT3 pathway inactivation in human colorectal cancer cells. Oncol. Lett. 9(4): Liu, J., S. Wang, Y. Zhang, H. Fan, and H. Lin Traditional Chinese medicine and cancer: history, present situation, and development. Thorac. Cancer. 6(5): Mols, F., T. Beijers, V. Lemmens, V. D. Hurk CJ, G. Vreugdenhil, V.D. Poll-Franse LV Chemotherapy-induced neuropathy and its association with quality of life among 2-to 11-year colorectal cancer survivors: results from the population-based PROFILES registry. J. Clin. Oncol. 31(21): Pan, X.Y., H. Guo, J. Han, F. Hao, Y. An, Y. Xu, Y. Xiaokaiti, Y. Pan, X.Li Ginsenoside Rg3 attenuates cell migration via inhibition of aquaporin 1 expression in PC-3M prostate cancer cells. Euro. J. Pharm. 683(1): Seo, E., and W. Kim Red ginseng extract reduced metastasis of colon cancer cells in vitro and in vivo. J. Ginseng. Res. 35(3): Shao, J., G. Gong, and L. Trombetta An evidence-based perspective of hedyotis diffusa or oldenlandia diffusa (spreading hedyotis) for cancer patients. In William C.S. Cho (Ed.), Evidence-Based Anticancer MateriaMedica. Springer. pp Wang, X., F. Zhang, L. Yang, Y. Mei, H. Long, X. Zhang, J. Zhang, Q. Suyila, X. Su Ursolic acid inhibits proliferation and induces apoptosis of cancer cells in vitro and in vivo. J. Biomed. Biotech doi: /2011/ Weidong, L., L. Cheng, Z. Honggang, C. Guohong, and H. Baojin Therapeutic targets of traditional Chinese medicine for colorectal cancer. J. Trad. Chin. Med. 36(2):

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