Effects of TOFA on growth of Eca109 and KYSE-450 cells in human esophageal squamous cell carcinoma

doi:10.12047/j.cjap.6236.2022.060

Abstract

Abstract: Objective: To investigate the effects of 5-tetradecanoxy 2-furanic acid (TOFA) on cell proliferation, cell cycle and apoptosis of esophageal squamous cell carcinoma (ESCC) cells. Methods: Eca-109 cells and KYSE-450 cells were divided into control group (DMSO) and experimental group (TOFA), respectively. The cells (4×10³ cells/100 μl) were inoculated into 96-well plates with 5 multiple wells at each concentration. After 24 h culture, cells were treated with DMSO or different concentrations (1, 3, 5, 10 μg/ ml) of TOFA for 24, 48 and 72 h. Cell proliferation was detected by MTT, cell cycle and apoptosis were detected by flow cytometry, the expression levels of p21 and Cleaved caspase-3 and modification levels of p-Akt, p-mTOR and p-4EBP1 were detected by Western blot, and intracellular free fatty acids were detected by special kits. Results: MTT results showed that TOFA inhibited the proliferation of Eca109 and KYSE-450 cells in a concentration and time dependent manner (all P<0.05), with IC₅₀ of 4.65 μg/ml and 3.93 μg/ml for 48 h, respectively. Flow cytometry results showed that compared with DMSO group, the percentage of cells in G2/M phase was increased and the apoptosis rate was increased in the experimental group. Western blotting results showed that compared with DMSO group, p21 and Cleaved caspase-3 protein expression levels were up-regulated, and p-AKT, p-mTOR and p-4EBP1 protein expression levels were down-regulated (all P<0.05). Conclusion: TOFA inhibits the proliferation, blocks the cycle progression and promotes apoptosis of ESCC, the mechanism may be related to the AKT/mTOR/4EBP1 signaling pathway.

Key words: esophageal squamous cell carcinoma, 5-tetradecanoxy 2-furanic acid (TOFA), proliferation, cycle, apoptosis, cell culture

CLC Number:

R73-3

GU Cheng-wei, QIAN He, LIU Yu-zhen, ZHAO Bao-sheng. Effects of TOFA on growth of Eca109 and KYSE-450 cells in human esophageal squamous cell carcinoma[J]. CJAP, 2022, 38(4): 317-321.

References

[1] 郑荣寿, 孙可欣, 赫捷, 等. 2015年中国恶性肿瘤流行情况分析[J]. 中华肿瘤杂志, 2019, 41(1): 19-28.
[2] Xu QL, Li H, Zhu YJ, et al. The treatments and postoperative complications of esophageal cancer: a review[J]. J Cardiothorac Surg, 2020, 15(1): 1-10.
[3] Wang C, Xu C, Sun M, et al. Acetyl-CoA carboxylase-alpha inhibitor TOFA induces humancancer cell apoptosis[J]. Biochem Biophys Res Commun, 2009, 385(3): 302-306.
[4] Guseva NV, Rokhlin OW, Glover RA, et al. TOFA (5-tetradecyl-oxy-2-furoic acid) reducesfatty acid synthesis, inhibits expression of AR, neuropilin-1 and Mcl-1 and kills prostate cancer cells independent of p53 status[J]. Cancer Biol Ther, 2011, 12(1): 80-85.
[5] Abbas G, Krasna M.Overview of esophageal cancer[J]. Ann Cardiothorac Surg, 2017, 6(2): 131-136.
[6] Pope ED 3rd, Kimbrough EO, Vemireddy LP, et al. Aberrant lipid metabolism as a therapeutic target in liver cancer[J]. Expert Opin Ther Targets, 2019, 23(6): 473-483.
[7] 金家豪, 赵宝生, 刘丹辉, 等. 低氧通过上调乙酰辅酶A羧化酶1促进肺腺癌A549细胞迁移[J]. 解剖学报, 2021, 52(2): 258-263.
[8] Monaco ME.Fatty acid metabolism in breast cancersubtypes[J]. Oncotarget, 2017, 8(17): 29487-29500.
[9] Menendez JA, Lupu R.Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis[J]. Nat Rev Cancer, 2007, 7(10): 763-777.
[10] Wang X, Tian J, Zhao Q, et al. Functional characterization of a low-frequency V1937I variant in FASN associated with susceptibility to esophageal squamous cellcarcinoma[J]. Arch Toxicol, 2020, 94(6): 2039-2046.
[11] He Y, Hua R, Li B, et al. Loss of FBP1 promotes proliferation, migration, and invasion by regulating fatty acid metabolism in esophageal squamous cell carcinoma[J]. Aging, 2020, 13(4): 4986-4998.
[12] Fan TJ, Han LH, Cong RS, et al. Caspase family proteases and apoptosis[J]. Acta Biochim Biophys Sin, 2005, 37(11): 719-727.
[13] Alzahrani AS.PI3K/AKT/mTOR inhibitors in cancer: At the bench and bedside[J]. Semin Cancer Biol, 2019, 59: 125-132.
[14] Zhou BP, Hung MC.Novel targets ofAkt, p21(Cipl/WAF1), and MDM2[J]. Semin Oncol, 2002, 29: 62-70.
[15] Jeong YJ, Hoe HS, Cho HJ, et al. Suppression of c-Myc enhances p21WAF1/CIP1-mediated G1 cell cycle arrest through the modulation of ERK phosphorylation byascochlorin[J]. J Cell Biochem, 2018, 119(2): 2036-2047.
[16] Kannan-Thulasiraman P, Dolniak B, Kaur S, et al. Role of the translational repressor 4E-BP1 in the regulation ofp21(Waf1/Cip1) expression by retinoids[J]. Biochem Biophys Res Commun, 2008, 368(4): 983-989.
[17] Zhou H, Li M, Meink J, et al. Akt regulates cell survival and apoptosis at a postmitochondriallevel[J]. J Cell Biol, 2000, 151(3): 483-494.
[18] 丁亦含, 李玉峰. mTOR 信号通路与自噬、凋亡之间的相互关系[J]. 现代医学, 2015, 43(6): 801-804.
[19] Hennessy BT, Smith DL, Ram PT, et al. Exploiting the PI3K/AKT pathway for cancer drug discovery[J]. Nat Rev Drug Discov, 2005, 4(12): 988-1004.