Human Colon Cancer Cell SW620 Research Articles

Design, synthesis, and antiproliferative activity of novel 1,2,4-triazole-chalcone compounds

Abstract A series of novel 1,2,4-triazole-chalcone compounds 10a–10s were designed by molecular hybridization strategy and synthesized. The molecular structures of the novel chalcones were characterized by 1H-NMR, 13C-NMR, and HRMS. The anti-proliferative activities of the novel chalcones against four cancer cell lines in vitro were examined by MTT, four tumor cell lines including human colon cancer cell SW620, human non-small cell lung cancer cell A549, human cervical cancer cell HeLa, and human breast cancer cell MCF-7. Compound 10o showed certain selectivity to SW620 cell line, and its IC50 value was 21.55 μM. 10q had good anti-proliferative activity against A549 cells with an IC50 value of 25.58 μM.

In vitro antioxidant, anti-inflammatory, and anticancer activities of mixture Thai medicinal plants

BackgroundThe phytochemical study of medicinal plants is rapidly gaining popularity with many pharmacologic effects. This study aims to determine the antioxidant capacity as well as anticancer and antimigration activities of Clear belongs Plus extract (CBL-P) which consisted of five medicinal plants namely, Alpinia galanga, Piper nigrum, Citrus aurantifolia, Tiliacora triandra, and Cannabis sativa on human colon cancer cells SW620 and HCT116 cell lines, and human non-small cell lung cancer cells A549 and NCI-H460 cell lines.MethodsIn this study the dried-plant powder was extracted using 90% ethanol. Additionally, CBL-P was studied antioxidative activity via DPPH and ABTS assays and anti-inflammatory activities using nitric oxide assay using Griess reaction. Antiproliferation and antimigration of CBL-P were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and scratch assay.ResultsThe results showed that CBL-P had potent antiproliferative activity with IC50 values in a concentration- and time-dependent manners for all four cell lines. CBL-P also possessed potent antimigration activity against all studied cancer cells. CBL-P demonstrated antimigration activity on four different types of cancer cells (A549, NCI-H460, HCT116, and SW620) after 48 h of incubation, with the greatest effect seen at the highest concentration tested (15 μg/mL) in A549 cells (10.23% of wound closure) and NCI-H460 cells (9.16% of wound closure). CBL-P was also effective in reducing migration in HCT116 and SW620 cells, with a range of closure area from 10—50%. In addition, CBL-P had antioxidant activity with IC50 values of 8.549 ± 0.241 mg/mL and 2.673 ± 0.437 mg/mL for DPPH and ABTS assays, respectively. CBL-P also showed anti-inflammatory activity with the best inhibitory activity on NO production at a concentration of 40 μg/mL.ConclusionIn conclusion, the mixture extract possessed antioxidant and anti-inflammatory activity. Furthermore, the mixture plant extract significantly exhibited antiproliferative and antimigration activities on SW620, HCT116, A549, and NCI-H460 cells (P ≤ 0.05). Taken together, our results suggest that medicinal plants may have synergistic effects that could potentially enhance the effectiveness of cancer treatment when used as adjuvants. These findings provide a solid scientific foundation for future efforts to explore the mechanism of action.

Abstract A83: In vitro and In vivo antitumor activity of immune suppressant FTY720

Abstract Purpose: Sphingosine-1-phosphate (S1P) is a pleotropic bioactive lipid mediator involved in the pathogenesis of inflammation and cancer. FTY720 is a “functional antagonist” of S1P by binding and induces the internalization and degradation of S1P receptors. Indeed, FTY720 was shown to have antitumor activity in several tumor models, while whether its immune-suppressive activity promotes or inhibits tumor growth is still unclear. In this study, we explored the antitumor activity of FTY720 in various cancer cells and its underlying mechanisms. Experimental Procedures: We analyzed the antiproliferative activity of FTY720 in nearly 30 human cancer cell lines and 6 mouse cancer cell lines. Human colon cancer cells SW620 and mouse breast cancer cells 4T1, which are sensitive to FTY720, were selected and used for in vivo study. SW620 was subcutaneously inoculated into Balb/c nude mice and 4T1 were in situ inoculated into immune-competent Balb/c mice. Mice was administrated intraperitoneally with low dose (0.5 mg/kg/d) and high dose (5 mg/kg/d) of FTY720. Tumor volume and body weight were measured every 3 days. After 3 weeks, the mice were sacrificed, and the tumor was removed and weighed, and kept for further analysis. The signaling pathway involved in S1P pathway and neovascularization were analyzed. In addition, the pharmacokinetics of FTY720 was also determined in mice. The effect of FTY720 on angiogenesis was also analyzed at cellular level, as well as the gene expression in HUVEC cells. Summary: FTY720 showed potent antiproliferative activity across various cancer cell lines. The IC50 for SW620 and 4T1 was 3.71 μM and 3.38 μM respectively, which were most sensitive. In animal study, both low-dose and high-dose FTY720 dramatically inhibited the tumor growth in SW620 xenograft model, while only high-dose FTY720 inhibited tumor growth in 4T1 xenograft model. Since the serum concentration of FTY720 was much lower than its cytotoxicity at low dose, we thus propose the anti-umor activity of FTY720 may due to its antiangiogenic activity. Moreover, the antitumor activity of FTY720 was much lower for 4T1 as compared to SW620 xenograft model, which possibly is caused by its immune-suppressive activity as it depleted the circulating lymphocytes. Besides, FTY720 dramatically inhibited neovascularization in tumor tissues. The half-life and AUClast are good for FTY720 in mice. FTY720 inhibited new vessel formation at cellular level. In addition, the microarray data showed the low-dose FTY720 (100 nM) induced the alteration of a few genes’ expression. Conclusion: FTY720 has potent antitumor activity both in vitro and in vivo, the mechanism of which is associated with proapoptotic, antiangiogenic and anti-inflammatory activities. The immune-suppressive activity of FTY720 may limit the antitumor activity in immune-competent individuals. This study provides a comprehensive understanding of the immune-suppressant FTY720 in antitumor. Note: This abstract was not presented at the conference. Citation Format: Zuoquan Xie, Xiaoxu Chen, Hui Xu, Jing Ai, Meiyu Geng. In vitro and In vivo antitumor activity of immune suppressant FTY720 [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A83.

Abstract B61: Transcriptomic landscape of the immune suppressant FTY720 in antitumor

Abstract Purpose: Sphingosine-1-phosphate (S1P) is a bioactive lipid that promotes cell survival and proliferation, and modulation of lymphocyte egress. It plays important roles in the pathogenesis of inflammation and cancer. FTY720 is a “functional antagonist” of S1P by binding and induces the internalization and degradation of S1P receptors. FTY720 showed antitumor activity in several tumor models, but the mechanism has not been fully illustrated. In this study, we used microarray to explore the signaling pathways and genes regulated by FTY720, which may help to illustrate the mechanism of action. Experimental Procedures: We tested the inhibitory activity of FTY720 in various human cancer cell lines. Human colon cancer cells SW620, which are sensitive to FTY720, were selected and used for further study. SW620 was subcutaneously inoculated into Balb/c nude mice, then treated intraperitoneally with low dose (0.5 mg/kg/d) and high dose (5 mg/kg/d) of FTY720. Meanwhile, microarray analysis was used to explore the modulated genes by FTY720. Moreover, we tested the anti-neovascularization effect of FTY720 on endothelial cells (HMEC), which was shown to have this activity in vivo, and we also profiled the differentially expressed genes regulated by FTY720 on HMEC cells. Summary: FTY720 showed potent inhibitory activity on various cancer cell lines. The colon cancer cell SW620 was one of the most sensitive cells in response to FTY720 with an IC50 of 3.71 μM. FTY720 also inhibited new vessel formation at cellular level. In animal study, both low-dose and high-dose FTY720 dramatically inhibited the tumor growth in SW620 xenograft model. Since the serum concentration of FTY720 was much lower than its cellular cytotoxicity at low dose, we propose the antitumor activity may not only be due to a direct effect on tumor growth, but also on neovascularization and lowering of circulating lymphocytes. In transcriptomic study, we found cell cycle and DNA repair pathways were downregulated by FTY720 both in SW620 and HMEC cells. In HMEC cells, mTORC1, MYC, P53, FOXO1, and immune response pathways were downregulated while steroid metabolism and terpenoid backbone biosynthesis pathways were upregulated by FTY720. In SW620 cells, fatty acid metabolism, FOXO1, inflammatory and immune response pathways were downregulated while steroid metabolism and terpenoid backbone biosynthesis pathways were upregulated by FTY720. Thus, there are many overlaps of regulated pathways and genes by FTY720 between SW620 and HMEC cells. Interestingly, the angiogenesis and blood vessel morphogenesis were both downregulated by FTY772 in SW620 and HMEC cells. Conclusion: FTY720 has potent antitumor activity in vitro and in vivo. It showed pleiotropic effects on gene expression, especially downregulating those genes involved in cell cycle, angiogenesis, and inflammatory and immune response. This study displayed a landscape of differentially expressed genes modulated by FTY720 and provided a basis for further mechanistic study. Citation Format: Zuoquan Xie, Xiaoxu Chen, Qingzhong Zeng, Meiyu Geng. Transcriptomic landscape of the immune suppressant FTY720 in antitumor [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr B61.

Experimental Study of Specific Targeted Inhibition from RNAi Silencing CXCR7 to Human Colon Cancer Cell SW620

Object: To investigate the change of CXCR7 protein expression after CXCR7-shRNA transfered into human colon cancer cell SW620 which is lentivirus-mediated. Methods: 1. To design and synthesis three shRNA sequence and one negative control sequence of CXCR7. To synthesis and construct recombinant lentiviral vector by pSilencerTM 4. 1 system and CXCR7. To transfer the vector into HEK293T cell for packaging viral and to detect titer. 2. To transfer the three different recombinant lentiviral vector and one negative control vector into human colon cancer cell SW620. To detect expression and silence efficiency of CXCR7 mRNA by RT-PCR. The group expressing the best silence efficiency of CXCR7-shRNA is selected as expression vector for next experiment. 3. To detect the change of SW620 cell proliferation after CXCR7-shRNA transfection by MTT. 4. To detect the change of SW620 cell invasion and migration after CXCR7-shRNA transfection by cell scratching experiment. 5. To detect the SW620 protein expression after CXCR7-shRNA transfection by Western blot. Results: 1. Packaging the three different recombinant lentiviral vector and one negative control vector is successful by sequencing and titer is 3. 16 × 108TU / ml,4. 27 × 108TU / ml,3. 93 × 108TU / ml,2. 95 × 108TU / ml respectively. 2. Expression of CXCR7 mRNA in three positive group is lower than negative control group after transfection( P 0. 05). The inhibition rate of CXCR7-shRNA-1 to CXCR7 is higher than the another group. 3. The tumor cell proliferation is reduced after CXCR7-shRNA-1 transfection into SW620 cell and there was significant difference comparing to control group( P 0. 05). 4. The migration index of control group and positive group are( 49. 92 ± 6. 41) %,( 29. 13 ± 5. 38) % respectively 24 hours after cell scratching experiment and it has statistical significance( P 0. 05). The migration index of control group and positive group are( 96. 52 ± 7. 44) %,( 72. 03 ± 8. 29) % respectively 48h hours after cell scratching experiment and it has statistical significance( P 0. 05). 5. Expression of CXCR7 is reduced after transfection into SW620 cell comparing to empty virus vector group and control group and it has statistical significance( P 0. 05). Conclusion: Expression of CXCR7 mRNA and CXCR7 protein is down-regulated effectively after CXCR7-shRNA recombinant lentiviral vector transfection into SW620 cell and proliferation and migration of tumor cell is inhibited effectively. It is a good foundation on next study of colon cancer RNAi therapy with lentiviral which target CXCR7 / CXCL12 biological axis. And it is a new direction for colon cancer gene therapy.

Antiproliferation and anti-migration induced by gypenosides in human colon cancer SW620 and esophageal cancer Eca-109 cells

Gypenosides (Gyp), found in Gynostemma pentaphyllum Makino, has attracted more attention owing to its wide bioactivities. However, the effects of Gyp on esophageal cancer cells and colon cancer cells are still unknown. The present study was to investigate the possible anti-proliferative and anti-migration activity of Gyp on human colon cancer cells SW620 and esophageal cancer cells Eca-109. Cell viability was evaluated using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell membrane integrity was evaluated using flow cytometry following propidium iodide staining. Apoptotic cell death was determined by nuclear 4'-6-diamidino-2-phenylindole staining. Generation of intracellular reactive oxygen species (ROS) and changes in mitochondrial membrane potential (Δψ m) was analyzed by flow cytometry using 2',7'-dichlorofluorescein-diacetate and rhodamine 123 staining, respectively. Wound healing assay was carried out to investigate Gyp-inhibited migration of SW620 and Eca-109 cells. The results indicated that Gyp inhibited cell proliferation and migration in SW620 and Eca-109 cells in dose- and time-dependent manner. Gyp elevated intracellular ROS level, decreased the Δψ m, and induced apoptotic morphology such as cell shrinkage and chromatin condensation, suggesting oxidative stress and mitochondria-dependent cell apoptosis that might be involved in Gyp-induced cell viability loss in SW620 and Eca-109 cells. The findings indicate Gyp may have valuable application in clinical colon cancer and esophageal cancer treatments.

Genistein reduced colon cancer metastasis to liver through regulating the expression of metastasis‐related genes in xenographed mice model

This study aimed at investigating the effect of genistein (Gen) on liver metastases derived from colon cancer. Male athymic nude mice were fed with western diet (W; calories%, fat 39%, carb 50%), W diet supplemented with 100ppm Gen (GL) or 500ppm Gen (GH). After 16 weeks, all the animals were intrasplenically injected with metastatic human colon cancer cells SW620 and sacrificed 8 weeks later. GL and GH mice showed lower liver metastases (36% and 17%) compared to W group (55% of mice with metastases). Mean size of metastases in the W group (4.7 mm2) was larger than that in GL (2.1 mm2) and GH (2.0 mm2) groups. To understand the molecular mechanism of the anti‐metastatic function of Gen, the expression of multiple metastasis‐related genes were determined by real‐time PCR. mRNA of NDRG1 gene in livers was greatly decreased by 50% and 60% in GL (p=0.005) and GH group (p=0.001) compared to W group. As NDRG1 has been reported as a biomarker for metastasis and poor prognosis in hepatocellular carcinoma, the down‐regulation of NDRG1 by Gen supplementation was corresponding to decreased liver metastasis. In summary, our study indicated that dietary supplementation of Gen suppressed colon cancer metastasis, which is associated with altered expression of metastasis‐related genes by the treatment.Grant Funding Source: university of illinois

Activation of several concurrent proapoptic pathways by sulforaphane in human colon cancer cells SW620

Despite the reported cytotoxicity and apoptosis-inducing properties of sulforaphane (SF) in colon cancer cells, the details concerning individual mechanisms and signaling cascades underlying SF-mediated apoptosis remain unclear. To understand different aspects of SF-induced proapoptic signaling in advanced colon carcinoma, we investigated its mechanisms in metastatic SW620 cell line. Our results indicate that in SW620 cells SF acts to induce multivariate cascades including DNA-damage response pathway whose proapoptotic signaling is nevertheless reduced owing to the mutant status of p53 and caspase-2-JNK pathway which seems to complement and enhance p53-dependent signaling, however only in wild-type p53. Furthermore, both pathways require the active role of mitochondria and do not depend on generation of ROS, making SF an attractive chemopreventive agent whose antitumor properties should be further investigated in colon cancer.