Mechanism of donafenib inhibiting proliferation, migration and invasion and promoting apoptosis of cholangiocarcinoma TFK-1 cells

doi:10.11958/20221485

Abstract

Abstract:

Objective To study the mechanism of donafenib inhibiting the proliferation, migration and invasion of human cholangiocarcinoma TFK-1 cells and promoting apoptosis. Methods Human cholangiocarcinoma TFK-1 cells were divided into the control group (treated with the same amount of dimethyl sulfoxide) and the 2, 5 and 10 μmol/L donafenib groups. CCK-8 assay was used to detect the inhibition rate of cell proliferation. Cell proliferation was detected by plate cloning assay. Cell apoptosis was detected by flow cytometry. Transwell assay was used to detect cell migration and invasion. Western blot assay was used to detect the expression of pathway proteins Wnt, β-catenin, Cyclin D1, anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax. Immunofluorescence assay was used to detect changes of β-catenin into the nucleus. Results With the increase of donafenib concentration, the proliferation inhibition rate and apoptosis rate of TFK-1 cells increased, the number of plate clone formation, cell migration and invasion decreased gradually, the expression levels of Wnt, β-catenin, Cyclin D1 and Bcl-2 decreased gradually, and the expression of Bax increased gradually (P<0.05). Immunofluorescence results showed that 2 μmol/L donafenib could inhibit β-catenin from entering the nucleus compared with the control group (P<0.05). Conclusion Donafenib can inhibit the proliferation, migration and invasion of human cholangiocarcinoma TFK-1 cells, and the mechanism may be related to the inhibition of Wnt/β-catenin pathway activation and the promotion of apoptosis.

Key words: bile duct neoplasms, Wnt signaling pathway, cell proliferation, cell movement, apoptosis, donafenib

CLC Number:

R735.8

Figures/Tables 7

References 19

[1]	GHOURI Y A, MIAN I, BLECHACZ B. Cancer review:Cholangiocarcinoma[J]. J Carcinog, 2015, 14:1. doi:10.4103/1477-3163.151940.
[2]	USON JUNIOR P, ARORA M, BOGENBERGER J M, et al. Recent advances in understanding cholangiocarcinoma[J]. Fac Rev, 2020, 9:15. doi:10.12703/b/9-15.
[3]	SHIMODA M, KUBOTA K. Multi-disciplinary treatment for cholangiocellular carcinoma[J]. World J Gastroenterol, 2007, 13(10):1500-1504. doi:10.3748/wjg.v13.i10.1500.
[4]	HUANG Y, LI X, ZHAO Y. Progression of targeted therapy in advanced cholangiocarcinoma[J]. Chin J Cancer Res, 2015, 27(2):122-127. doi:10.3978/j.issn.1000-9604.2015.04.01.
[5]	KEAM S J, DUGGAN S. Donafenib:First approval[J]. Drugs, 2021, 81(16):1915-1920. doi:10.1007/s40265-021-01603-0.
[6]	LI Q, ZHU H. Donafenib treatment for hepatocellular carcinoma:A case report[J]. Medicine(Baltimore), 2021, 100(25):e26373. doi:10.1097/MD.0000000000026373.
[7]	LIN Y S, YANG H, DING Y, et al. Donafenib in progressive locally advanced or metastatic radioactive iodine-refractory differentiated thyroid cancer:Results of a randomized,multicenter phase II trial[J]. Thyroid, 2021, 31(4):607-615. doi:10.1089/thy.2020.0235.
[8]	YANG Y, ZHU H, LI Q. Partial response of donafenib as the third-line therapy in metastatic colon cancer:A case report[J]. Medicine(Baltimore), 2021, 100(37):e27204. doi:10.1097/MD.0000000000027204.
[9]	SUKSEN K, JANPIPATKUL K, REABROI S, et al. Gambogic acid inhibits Wnt/β-catenin signaling and induces ER stress-mediated apoptosis in human cholangiocarcinoma[J]. Asian Pac J Cancer Prev, 2021, 22(6):1913-1920. doi:10.31557/APJCP.2021.22.6.1913.
[10]	ZHANG Y, WANG X. Targeting the Wnt/β-catenin signaling pathway in cancer[J]. J Hematol Oncol, 2020, 13(1):165. doi:10.1186/s13045-020-00990-3.
[11]	TIAN W, HU W, SHI X, et al. Comprehensive genomic profile of cholangiocarcinomas in China[J]. Oncol Lett, 2020, 19(4):3101-3110. doi:10.3892/ol.2020.11429.
[12]	MAEMURA K, NATSUGOE S, TAKAO S. Molecular mechanism of cholangiocarcinoma carcinogenesis[J]. J Hepatobiliary Pancreat Sci, 2014, 21(10):754-760. doi:10.1002/jhbp.126.
[13]	QIN S, BI F, GU S, et al. Donafenib versus sorafenib in first-line treatment of unresectable or metastatic hepatocellular carcinoma:A randomized,open-label,parallel-controlled phase II-III trial[J]. J Clin Oncol, 2021, 39(27):3002-3011. doi:10.1200/JCO.21.00163.
[14]	LIU J, XIAO Q, XIAO J, et al. Wnt/β-catenin signalling:Function,biological mechanisms,and therapeutic opportunities[J]. Signal Transduct Target Ther, 2022, 7(1):3. doi:10.1038/s41392-021-00762-6.
[15]	PERUGORRIA M J, OLAIZOLA P, LABIANO I, et al. Wnt-β-catenin signalling in liver development, health and disease[J]. Nat Rev Gastroenterol Hepatol, 2019, 16(2):121-136. doi:10.1038/s41575-018-0075-9.
[16]	ZHANG G F, QIU L, YANG S L, et al. Wnt/β-catenin signaling as an emerging potential key pharmacological target in cholangiocarcinoma[J]. Biosci Rep, 2020, 40(3):BSR20193353. doi:10.1042/BSR20193353.
[17]	HANAHAN D. Hallmarks of cancer:New dimensions[J]. Cancer Discov, 2022, 12(1):31-46. doi:10.1158/2159-8290.CD-21-1059.
[18]	YANG X, WANG S, MU Y, et al. Schisandrin B inhibits cell proliferation and induces apoptosis in human cholangiocarcinoma cells[J]. Oncol Rep, 2016, 36(4):1799-1806. doi:10.3892/or.2016.4992.
[19]	LI V S, NG S S, BOERSEMA P J, et al. Wnt signaling through inhibition of β-catenin degradation in an intact Axin1 complex[J]. Cell, 2012, 149(6):1245-1256. doi:10.1016/j.cell.2012.05.002.

组别	n	细胞迁移	细胞侵袭
对照组	3	167.67±5.31	216.67±4.64
2 μmol/L多纳非尼组	3	77.33±4.50^a	116.33±3.40^a
5 μmol/L多纳非尼组	3	40.67±2.87^ab	33.67±2.49^ab
10 μmol/L多纳非尼组	3	27.67±2.49^abc	19.33±2.62^abc
F		507.400^＊＊	1 427.000^＊＊

组别	n	细胞迁移	细胞侵袭
对照组	3	167.67±5.31	216.67±4.64
2 μmol/L多纳非尼组	3	77.33±4.50^a	116.33±3.40^a
5 μmol/L多纳非尼组	3	40.67±2.87^ab	33.67±2.49^ab
10 μmol/L多纳非尼组	3	27.67±2.49^abc	19.33±2.62^abc
F		507.400^＊＊	1 427.000^＊＊

组别	Wnt	β-catenin	Cyclin D1	Bax	Bcl-2
对照组	0.72±0.05	0.69±0.03	0.80±0.04	0.15±0.03	0.53±0.02
2 μmol/L多纳非尼组	0.53±0.02^a	0.46±0.03^a	0.63±0.03^a	0.27±0.02^a	0.32±0.02^a
5 μmol/L多纳非尼组	0.33±0.04^ab	0.34±0.02^ab	0.34±0.02^ab	0.42±0.01^ab	0.23±0.02^ab
10 μmol/L多纳非尼组	0.17±0.04^abc	0.18±0.01^abc	0.31±0.02^abc	0.90±0.03^abc	0.13±0.03^abc
F	514.400^＊＊	438.000^＊＊	594.300^＊＊	990.800^＊＊	475.000^＊＊

组别	Wnt	β-catenin	Cyclin D1	Bax	Bcl-2
对照组	0.72±0.05	0.69±0.03	0.80±0.04	0.15±0.03	0.53±0.02
2 μmol/L多纳非尼组	0.53±0.02^a	0.46±0.03^a	0.63±0.03^a	0.27±0.02^a	0.32±0.02^a
5 μmol/L多纳非尼组	0.33±0.04^ab	0.34±0.02^ab	0.34±0.02^ab	0.42±0.01^ab	0.23±0.02^ab
10 μmol/L多纳非尼组	0.17±0.04^abc	0.18±0.01^abc	0.31±0.02^abc	0.90±0.03^abc	0.13±0.03^abc
F	514.400^＊＊	438.000^＊＊	594.300^＊＊	990.800^＊＊	475.000^＊＊