Iguratimod enhances mitomycin C-induced apoptosis in human esophageal cancer Eca109 cells

doi:10.3877/cma.j.issn.2095-1221.2019.01.003

Abstract

Abstract:

Objective

To investigate the effect of the combination of Iguratimod (T-?614) and mitomycin (MMC) on apoptosis of human esophageal cancer Eca109 cells, and to explore the possible mechanism.

Methods

Eca109 cells were exposed to MMC in the presence or absence of difference concentrations of T-614. Eca109 cell viability and reactive oxygen species (ROS) production were detected by CCK8 and DCFH-DA staining. Apoptosis of Eca109 cells was detected by flow cytometry. Expression of TNFα and cleaved caspase3 in Eca109 cells was detected by Western Blot. Statistical analysis was performed using one-way analysis of variance and t test.

Results

CCK8 results showed that A450 values of 25, 50, and 100 μg/ml T-614+MMC group (0.73±0.02, 0.52±0.02, and 0.33±0.02) were significantly reduced compared with A450 value (0.85±0.03) of 0 μg/?ml T-614+MMC group (F?=?127.60, P?< 0.01) . The results of DCFH-DA staining showed that the DCFH values of the 25, 50, and 100 μg/ml T-614+MMC group (219.67±9.50, 280.33±10.50, and 338.33±16.07) were significantly increased compared with the fluorescence value of DCFH in the 0?μg/ml T-614+MMC group (130.00±10.00) . The difference was statistically significant (F?=?170.20, P?<?0.01) . The results of flow cytometry showed that the apoptosis rate of T-614 group (20.30±2.00) ﹪ and MMC group (25.60±3.00) ﹪ was significantly increased compared with the control group (5.33±0.35) ﹪. Compared with MMC group (25.60±3.00) ﹪, the apoptosis rate (56.20±3.00) ﹪of the T-614+MMC group was significantly increased (F?= 247.50, P?< 0.01) . Western Blot results showed that the expression of TNFα and cleaved caspase3 in T-614+MMC group (1.28±0.08, 0.59±0.03) was significantly increased compared with the expression in MMC group (0.87±0.06, 0.25±0.03) (F?= 96.90, P?< 0.01) .

Conclusion

T-614 significantly enhanced the inhibitory effect of MMC on the viability of Eca109 cells. The mechanism may be related to the production of ROS and the activation of mitochondrial apoptosis pathway, which ultimately leads to apoptosis of esophageal cancer Eca109 cells.

Key words: Esophageal cancer, Iguratimod, Reactive oxygen species, Apoptosis, Mitomycin

Yu Wu, Jialin Wang. Iguratimod enhances mitomycin C-induced apoptosis in human esophageal cancer Eca109 cells[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2019, 09(01): 13-17.

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References 21

[1]	Sabra MJ,Smotherman C,Kraemer DF, et al. The effects of neoadjuvant therapy on morbidity and mortality of esophagectomy for esophageal cancer: American college of surgeons National surgical quality improvement program (ACS-NSQIP) 2005-2012[J]. J Surg Oncol, 2017, 115(3):296-300.
[2]	Xi M,Liao ZX,Deng WY, et al. Recursive partitioning analysis identifies pretreatment risk groups for the utility of induction chemotherapy before definitive chemoradiation therapy in esophageal cancer[J]. Int J Radiat Oncol Biol Phys, 2017, 99(2):407-416.
[3]	Altorki N,Harrison S. What is the role of neoadjuvant chemotherapy, radiation, and adjuvant treatment in resectable esophageal cancer?[J]. Ann Cardiothorac Surg, 2017, 6(2):167-174.
[4]	Sun Y,Ye DW,Zhang P, et al. Anti-rheumatic drug iguratimod(T-614)alleviates cancer-induced bone destruction via down-regulating interleukin-6 production in a nuclear factor-kappaB-dependent manner[J]. J Huazhong Univ Sci Technolog Med Sci, 2016, 36(5):691-699.
[5]	Wang ZL,Zhou ZG,Chen Y, et al. Support vector machines model of computed tomography for assessing lymph node metastasis in esophageal cancer with neoadjuvant chemotherapy[J]. J Comput Assist Tomogr, 2017, 41(3):455-460.
[6]	Ku GY. Systemic therapy for esophageal cancer: chemotherapy[J]. Chin Clin Oncol, 2017, 6(5):49.
[7]	Hikami S,Shiozaki A,Fujiwara H, et al. [Outcome of Patients with Pathological Complete Response after Neoadjuvant Chemotherapy in Advanced Esophageal Cancer][J]. Gan To Kagaku Ryoho, 2017, 44(12):1796-1798.
[8]	Fujishima H,Fumoto S,Shibata T, et al. A 17-molecule set as a predictor of complete response to neoadjuvant chemotherapy with docetaxel, cisplatin, and 5-fluorouracil in esophageal cancer[J]. PLoS One, 2017, 12(11):e0188098.
[9]	Hara M,Abe T,Sugawara S, et al. Long-term safety study of iguratimod in patients with rheumatoid arthritis[J]. Mod Rheumatol, 2007, 17(1):10-16.
[10]	Liu D,Liu CF,Wang N, et al. The research of effects of iguratimod (T- 614) on the apoptosis of peripheral blood mononuclear cell and TH1 in rheumatoid arthritis[J]. Value Health, 2014, 17(7):A772.
[11]	Chen L,Xiong YQ,Xu J, et al. Juglanin inhibits lung cancer by regulation of apoptosis, ROS and autophagy induction[J]. Oncotarget, 2017, 8(55):93878-93898.
[12]	Wang Y,Yu XY,Song HT, et al. The STAT-ROS cycle extends IFN-induced cancer cell apoptosis[J]. Int J Oncol, 2018, 52(1):305-313.
[13]	Cho HD,Lee JH,Moon KD, et al. Auriculasin-induced ROS causes prostate cancer cell death via induction of apoptosis[J]. Food Chem Toxicol, 2018, 111:660-669.
[14]	Khan F,Khan I,Farooqui A, et al. Carvacrol induces reactive Oxygen species(ROS)-mediated apoptosis along with cell cycle arrest at G0/G1 in human prostate cancer cells[J]. Nutr Cancer, 2017, 69(7):1075-1087.
[15]	Khafaga AF,El-Sayed YS. All-trans-retinoic acid ameliorates doxorubicin-induced cardiotoxicity: in vivo potential involvement of oxidative stress, inflammation, and apoptosis via caspase-3 and p53 down-expression[J]. Naunyn Schmiedebergs Arch Pharmacol, 2018, 391(1):59-70.
[16]	Dyari HRE,Rawling T,Chen Y, et al. A novel synthetic analogue of omega-3 17,18-epoxyeicosatetraenoic acid activates TNF receptor-1/ASK1/JNK signaling to promote apoptosis in human breast cancer cells[J]. FASEB J, 2017, 31(12):5246.
[17]	Templin AT,Samarasekera T,Meier DT, et al. Apoptosis repressor with caspase recruitment domain ameliorates Amyloid-Induced beta-Cell apoptosis and JNK pathway activation[J]. Diabetes, 2017, 66(10):2636-2645.
[18]	Mohr A,Deedigan L,Jencz S, et al. Caspase-10: a molecular Switch from cell-autonomous apoptosis to communal cell death in response to chemotherapeutic drug treatment[J]. Cell Death Differ, 2018, 25(2):340-352.
[19]	Zhou PP,Wang CL,Hu ZB, et al. Genistein induces apoptosis of colon cancer cells by reversal of epithelial-to-mesenchymal via a Notchl/NF-kappa B/slug/E-cadherin pathway[J]. BMC Cancer, 2017, 17(1):813.
[20]	Zhao Y,Tian B,Wang Y, et al. Kaempferol Sensitizes Human Ovarian Cancer Cells-OVCAR-3 and SKOV-3 to Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL)-Induced Apoptosis via JNK/ERK-CHOP Pathway and Up-Regulation of Death Receptors 4 and 5 [J]. Med Sci Monit,2017, 23:5096-5105.
[21]	Thapa B,Bahadur RK,Uludag H. Novel targets for sensitizing breast cancer cells to TRAIL-induced apoptosis with siRNA delivery[J]. Int J Cancer, 2018, 142(3):597-606.

T-614分组浓度	样本数	T-614（A450）	T-614+MMC（A450）
0 μg/ml	3	1.05±0.05	0.85±0.03
25 μg/ml	3	0.98±0.08	0.73±0.02^a
50 μg/ml	3	0.82±0.08^a	0.52±0.02^a
100 μg/ml	3	0.55±0.05^a	0.33±0.02^a
F值	?	35.73	127.60
P值	?	< 0.0001	< 0.0001

T-614分组浓度	样本数	T-614（A450）	T-614+MMC（A450）
0 μg/ml	3	1.05±0.05	0.85±0.03
25 μg/ml	3	0.98±0.08	0.73±0.02^a
50 μg/ml	3	0.82±0.08^a	0.52±0.02^a
100 μg/ml	3	0.55±0.05^a	0.33±0.02^a
F值	?	35.73	127.60
P值	?	< 0.0001	< 0.0001

T-614分组浓度	样本数	T-614（DCFH）	T-614+MMC（DCFH）
0 μg/ml	3	35.00± 5.00	130.00±10.00
25 μg/ml	3	41.67± 1.53	219.67± 9.50^a
50 μg/ml	3	90.00±10.00^a	280.33±10.50^a
100 μg/ml	3	136.67±15.28^a	338.33±16.07^a
F值	?	74.71	170.20
P值	?	< 0.0001	< 0.0001

T-614分组浓度	样本数	T-614（DCFH）	T-614+MMC（DCFH）
0 μg/ml	3	35.00± 5.00	130.00±10.00
25 μg/ml	3	41.67± 1.53	219.67± 9.50^a
50 μg/ml	3	90.00±10.00^a	280.33±10.50^a
100 μg/ml	3	136.67±15.28^a	338.33±16.07^a
F值	?	74.71	170.20
P值	?	< 0.0001	< 0.0001

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