Experimental research of DAB2IP in regulating proliferation effect of colon cancer cells by salinomycin_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

SHEN Jie , JIA Weiyi , HUANG Qingwen , FENG Shengjie ,  LIU Liang

Department of Gastrointestinal Surgery & Gastrointestinal Cancer Research Institute, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P. R. China;

Corresponding author：

LIU Liang, Email: liangliu@tjh.tjmu.edu.cn

Keywords：

colon cancer; DAB2IP; salinomycin; cancer stem cell

DOI：

10.7507/1007-9424.202111014

Video：

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Objective To investigate the regulatory role of DAB2IP in proliferation effect of colon cancer cells by salinomycin. Methods Cell counting kit 8 (CCK8) assay was used to detect median inhibitory concentration (IC₅₀) of salinomycin on HT29 and SW480 cells. Colon cancer cells with stable knock-down of DAB2IP (HT29-shDAB2IP) and control cells (HT29-shcon) were constructed by lentivirus plasmid. And colon cancer cells with stable over-expression of DAB2IP (SW480-DAB2IP) and control cells (SW480-con) were constructed using pCI plasmid. The proliferation effect of salinomycin on stable knock-down or over-expression of DAB2IP by CCK8 assay in the colon cancer cell was identified. The colon cancer stem cells makers CD44, CD24, and CD133 were investigated using real-time PCR. Results The salinomycin had obvious inhibitory effects on the proliferations of HT29 and SW480 cells, the IC₅₀ value was 20.0 μmol/L and 10.0 μmol/L, respectively. The stable knock-down of DAB2IP could significantly enhance the inhibitory effect of salinomycin on the proliferation in HT29 cells (P<0.05) and stable over-expression of DAB2IP could significantly decrease the inhibitory effect of salinomycin on the proliferation in SW480 cells (P<0.05). Further the result of real-time PCR detection showed that the expressions of cancer stem cells markers CD44, D24, and CD133 were significantly increased after stable knock-down of DAB2IP in the HT29 cells (P<0.05), while the expressions were significantly decreased after stable over-expression of DAB2IP in the SW480 cells (P<0.05). Conclusions From initial results of this study, salinomycin could suppress proliferation of colon cancer cells. DAB2IP might weaken proliferative inhibitory effect of salinomycin by inhibiting expressions of cancer cells stem in colon cancer cells.

Citation： SHEN Jie, JIA Weiyi, HUANG Qingwen, FENG Shengjie, LIU Liang. Experimental research of DAB2IP in regulating proliferation effect of colon cancer cells by salinomycin. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2022, 29(7): 857-861. doi: 10.7507/1007-9424.202111014 Copy

1.	中华人民共和国国家卫生健康委员会医政医管局, 中华医学会肿瘤学分会. 中国结直肠癌诊疗规范(2020年版). 中国实用外科杂志, 2020, 40(6): 600-630.
2.	Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2018, 68(6): 394-424.
3.	Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med, 1997, 3(7): 730-737.
4.	Al-Hajj M, Wicha MS, Benito-Hernandez A, et al. Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA, 2003, 100(7): 3983-3988.
5.	Dalerba P, Dylla SJ, Park IK, et al. Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci USA, 2007, 104(24): 10158-10163.
6.	Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Res, 2003, 63(18): 5821-5828.
7.	Zhang H, Zhang R, Luo Y, et al. AIP1/DAB2IP, a novel member of the Ras-GAP family, transduces TRAF2-induced ASK1-JNK activation. J Biol Chem, 2004, 279(43): 44955-44965.
8.	Min W, Lin Y, Tang S, et al. AIP1 recruits phosphatase PP2A to ASK1 in tumor necrosis factor-induced ASK1-JNK activation. Circ Res, 2008, 102(7): 840-848.
9.	Xie D, Gore C, Liu J, et al. Role of DAB2IP in modulating epithelial-to-mesenchymal transition and prostate cancer metastasis. Proc Natl Acad Sci USA, 2010, 107(6): 2485-2490.
10.	Yun EJ, Baek ST, Xie D, et al. DAB2IP regulates cancer stem cell phenotypes through modulating stem cell factor receptor and ZEB1. Oncogene, 2015, 34(21): 2741-2752.
11.	Min J, Liu L, Li X, et al. Absence of DAB2IP promotes cancer stem cell like signatures and indicates poor survival outcome in colorectal cancer. Sci Rep, 2015, 5: 16578. doi: https://doi.org/10.1038/srep16578.
12.	Arafat K, Iratni R, Takahashi T, et al. Inhibitory effects of salinomycin on cell survival, colony growth, migration, and invasion of human non-small cell lung cancer A549 and LNM35: involvement of NAG-1. PLoS One, 2013, 8(6): e66931. doi: 10.1371/journal.pone.0066931.
13.	Lu Y, Ma W, Mao J, et al. Salinomycin exerts anticancer effects on human breast carcinoma MCF-7 cancer stem cells via modulation of Hedgehog signaling. Chem Biol Interact, 2015, 228: 100-107.
14.	Zhang Z, Zhao J, Mi Z, et al. Effects of salinomycin and 17-AAG on proliferation of human gastric cancer cells in vitro. Mol Med Rep, 2017, 16(2): 1063-1070.
15.	Zhou J, Li P, Xue X, et al. Salinomycin induces apoptosis in cisplatin-resistant colorectal cancer cells by accumulation of reactive oxygen species. Toxicol Lett, 2013, 222(2): 139-145.
16.	Parajuli B, Shin SJ, Kwon SH, et al. Salinomycin induces apoptosis via death receptor-5 up-regulation in cisplatin-resistant ovarian cancer cells. Anticancer Res, 2013, 33(4): 1457-1462.
17.	Klose J, Guerlevik E, Trostel T, et al. Salinomycin inhibits cholangiocarcinoma growth by inhibition of autophagic flux. Oncotarget, 2017, 9(3): 3619-3630.
18.	Sun J, Luo Q, Liu L, et al. Salinomycin attenuates liver cancer stem cell motility by enhancing cell stiffness and increasing F-actin formation via the FAK-ERK1/2 signalling pathway. Toxicology, 2017, 384: 1-10.
19.	Gupta PB, Onder TT, Jiang G, et al. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell, 2009, 138(4): 645-659.
20.	Mai TT, Hamaï A, Hienzsch A, et al. Salinomycin kills cancer stem cells by sequestering iron in lysosomes. Nat Chem, 2017, 9(10): 1025-1033.
21.	Dong TT, Zhou HM, Wang LL, et al. Salinomycin selectively targets ‘CD133⁺’ cell subpopulations and decreases malignant traits in colorectal cancer lines. Ann Surg Oncol, 2011, 18(6): 1797-1804.
22.	Wang Y. Effects of salinomycin on cancer stem cell in human lung adenocarcinoma A549 cells. Med Chem, 2011, 7(2): 106-111.
23.	Zhi QM, Chen XH, Ji J, et al. Salinomycin can effectively kill ALDH (high) stem-like cells on gastric cancer. Biomed Pharmacother, 2011, 65(7): 509-515.
24.	Tang QL, Zhao ZQ, Li JC, et al. Salinomycin inhibits osteosarcoma by targeting its tumor stem cells. Cancer Lett, 2011, 311(1): 113-121.
25.	Magrath JW, Kim Y. Salinomycin’s potential to eliminate glioblastoma stem cells and treat glioblastoma multiforme (Review). Int J Oncol, 2017, 51(3): 753-759.
26.	Munro MJ, Wickremesekera SK, Peng L, et al. Cancer stem cells in colorectal cancer: a review. J Clin Pathol, 2018, 71(2): 110-116.

1. 中华人民共和国国家卫生健康委员会医政医管局, 中华医学会肿瘤学分会. 中国结直肠癌诊疗规范(2020年版). 中国实用外科杂志, 2020, 40(6): 600-630.
2. Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2018, 68(6): 394-424.
3. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med, 1997, 3(7): 730-737.
4. Al-Hajj M, Wicha MS, Benito-Hernandez A, et al. Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA, 2003, 100(7): 3983-3988.
5. Dalerba P, Dylla SJ, Park IK, et al. Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci USA, 2007, 104(24): 10158-10163.
6. Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Res, 2003, 63(18): 5821-5828.
7. Zhang H, Zhang R, Luo Y, et al. AIP1/DAB2IP, a novel member of the Ras-GAP family, transduces TRAF2-induced ASK1-JNK activation. J Biol Chem, 2004, 279(43): 44955-44965.
8. Min W, Lin Y, Tang S, et al. AIP1 recruits phosphatase PP2A to ASK1 in tumor necrosis factor-induced ASK1-JNK activation. Circ Res, 2008, 102(7): 840-848.
9. Xie D, Gore C, Liu J, et al. Role of DAB2IP in modulating epithelial-to-mesenchymal transition and prostate cancer metastasis. Proc Natl Acad Sci USA, 2010, 107(6): 2485-2490.
10. Yun EJ, Baek ST, Xie D, et al. DAB2IP regulates cancer stem cell phenotypes through modulating stem cell factor receptor and ZEB1. Oncogene, 2015, 34(21): 2741-2752.
11. Min J, Liu L, Li X, et al. Absence of DAB2IP promotes cancer stem cell like signatures and indicates poor survival outcome in colorectal cancer. Sci Rep, 2015, 5: 16578. doi: https://doi.org/10.1038/srep16578.
12. Arafat K, Iratni R, Takahashi T, et al. Inhibitory effects of salinomycin on cell survival, colony growth, migration, and invasion of human non-small cell lung cancer A549 and LNM35: involvement of NAG-1. PLoS One, 2013, 8(6): e66931. doi: 10.1371/journal.pone.0066931.
13. Lu Y, Ma W, Mao J, et al. Salinomycin exerts anticancer effects on human breast carcinoma MCF-7 cancer stem cells via modulation of Hedgehog signaling. Chem Biol Interact, 2015, 228: 100-107.
14. Zhang Z, Zhao J, Mi Z, et al. Effects of salinomycin and 17-AAG on proliferation of human gastric cancer cells in vitro. Mol Med Rep, 2017, 16(2): 1063-1070.
15. Zhou J, Li P, Xue X, et al. Salinomycin induces apoptosis in cisplatin-resistant colorectal cancer cells by accumulation of reactive oxygen species. Toxicol Lett, 2013, 222(2): 139-145.
16. Parajuli B, Shin SJ, Kwon SH, et al. Salinomycin induces apoptosis via death receptor-5 up-regulation in cisplatin-resistant ovarian cancer cells. Anticancer Res, 2013, 33(4): 1457-1462.
17. Klose J, Guerlevik E, Trostel T, et al. Salinomycin inhibits cholangiocarcinoma growth by inhibition of autophagic flux. Oncotarget, 2017, 9(3): 3619-3630.
18. Sun J, Luo Q, Liu L, et al. Salinomycin attenuates liver cancer stem cell motility by enhancing cell stiffness and increasing F-actin formation via the FAK-ERK1/2 signalling pathway. Toxicology, 2017, 384: 1-10.
19. Gupta PB, Onder TT, Jiang G, et al. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell, 2009, 138(4): 645-659.
20. Mai TT, Hamaï A, Hienzsch A, et al. Salinomycin kills cancer stem cells by sequestering iron in lysosomes. Nat Chem, 2017, 9(10): 1025-1033.
21. Dong TT, Zhou HM, Wang LL, et al. Salinomycin selectively targets ‘CD133⁺’ cell subpopulations and decreases malignant traits in colorectal cancer lines. Ann Surg Oncol, 2011, 18(6): 1797-1804.
22. Wang Y. Effects of salinomycin on cancer stem cell in human lung adenocarcinoma A549 cells. Med Chem, 2011, 7(2): 106-111.
23. Zhi QM, Chen XH, Ji J, et al. Salinomycin can effectively kill ALDH (high) stem-like cells on gastric cancer. Biomed Pharmacother, 2011, 65(7): 509-515.
24. Tang QL, Zhao ZQ, Li JC, et al. Salinomycin inhibits osteosarcoma by targeting its tumor stem cells. Cancer Lett, 2011, 311(1): 113-121.
25. Magrath JW, Kim Y. Salinomycin’s potential to eliminate glioblastoma stem cells and treat glioblastoma multiforme (Review). Int J Oncol, 2017, 51(3): 753-759.
26. Munro MJ, Wickremesekera SK, Peng L, et al. Cancer stem cells in colorectal cancer: a review. J Clin Pathol, 2018, 71(2): 110-116.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Experimental research of DAB2IP in regulating proliferation effect of colon cancer cells by salinomycin

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