ALKBH5在甲状腺癌细胞中的生物学功能

doi:10.3969/j.issn.1004-616x.2024.05.003

Abstract

Abstract: OBJECTIVE： To investigate the role of N⁶-methyladenosine (m⁶A) demethylase ALKBH5 on intracellular and cellular functions of thyroid carcinoma cells in vitro. METHODS： The ALKBH5 knockdown plasmid was transfected to human thyroid papillary carcinoma cells (TPC-1) and undifferentiated thyroid carcinoma cells (8505C). Expression of m⁶A in the cancer cells were measured using a m⁶A methylation quantification detection kit. Plate cloning，CCK-8，plate scratch，and Transwell were used to analyze the proliferation，migration，and invasion ability. Flow cytometry was used to detect the cell cycle of the cancer cells and the percentage of cell in each phase. The relationship between ALKBH5 and downstream pathways was analyzed by Western blot experiments. RESULTS： Compared to cells without the ALKBH5 knockdown，the knockdown increased m⁶A expression of levels in the TPC-1 and 8505C cancer cells，and decreased the number of cell proliferation，invasion，and migration decreased (P<0.05). In addition，cells were arrested at the G₂/M phase of cell cycle. Western blot experiments showed that when the expression of ALKBH5 was low，the key genes of the PI3K/AKT signaling pathway were perturbed. Protein expressions of P85，AK，P-AKT and FAK were inhibited，as well as that of CDK4，CDK6，Cyclin B1，Cyclin D1，Cyclin E1 in the downstream Cyclin pathway，but P53 was increased (P<0.05). CONCLUSION： In the thyroid papillary carcinoma cells，ALKBH5 knockdown inhibited their proliferation，migration，invasion and cell cycle，as well as the PI3K/AKT/P53 and PI3K/AKT/Cyclin axis.

Key words: m⁶A, ALKBH5, thyroid carcinoma, PI3K/AKT signaling pathway

CLC Number:

WANG Shujun, ZHANG Huixia, WEN Sini, ZHANG Lei, WANG Xin, WU Anqi, ZHU Xiaonian, ZHANG Xiaoying, TAN Shengkui, HAN Fei. Intracellular and cellular functions of ALKBH5 in thyroid cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(5): 349-358.

References

[1] SUNG H, FERLAY J, SIEGEL R L, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249.
[2] MIRANDA-FILHO A, LORTET-TIEULENT J, BRAY F, et al. Thyroid cancer incidence trends by histology in 25 countries: a population-based study[J]. Lancet Diabetes Endocrinol, 2021, 9(4): 225-234.
[3] WANG K J, JIANG L, ZHANG Y, et al. Progression of thyroid carcinoma is promoted by the m⁶A methyltransferase mettl3 through regulating m⁶A methylation on TCF1[J]. Onco Targets Ther, 2020, 13: 1605-1612.
[4] OERUM S, MEYNIER V, CATALA M, et al. A comprehensive review of m⁶A/m⁶Am RNA methyltransferase structures[J]. Nucleic Acids Res, 2021, 49(13): 7239-7255.
[5] 钱晓芬, 曾平, 刘金富, 等. m⁶A RNA甲基化修饰相关酶的研究进展[J]. 中国免疫学杂志, 2023, 39(5): 1073-1084.
[6] YANG Y, HSU P J, CHEN Y S, et al. Dynamic transcriptomic m⁶A decoration: writers, erasers, readers and functions in RNA metabolism[J]. Cell Res, 2018, 28(6): 616-624.
[7] ZHENG G Q, DAHL J A, NIU Y M, et al. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility[J]. Mol Cell, 2013, 49(1): 18-29.
[8] CHEN W Z, ZHANG L, ZHENG G Q, et al. Crystal structure of the RNA demethylase ALKBH5 from zebrafish[J]. FEBS Lett, 2014, 588(6): 892-898.
[9] XU C, LIU K, TEMPEL W, et al. Structures of human ALKBH5 demethylase reveal a unique binding mode for specific single-stranded N⁶-methyladenosine RNA demethylation[J]. J Biol Chem, 2014, 289(25): 17299-17311.
[10] ZHU Y, PENG X Z, ZHOU Q L, et al. METTL3-mediated m⁶A modification of STEAP2 mRNA inhibits papillary thyroid cancer progress by blocking the Hedgehog signaling pathway and epithelial-to-mesenchymal transition[J]. Cell Death Dis, 2022, 13(4): 358.
[11] YANG X, ZHANG S, HE C Y, et al. METTL14 suppresses proliferation and metastasis of colorectal cancer by down-regulating oncogenic long non-coding RNA XIST[J]. Mol Cancer, 2020, 19(1): 46.
[12] WANG M, LIU J, ZHAO Y, et al. Upregulation of METTL14 mediates the elevation of PERP mRNA N⁶ adenosine methylation promoting the growth and metastasis of pancreatic cancer[J]. Mol Cancer, 2020, 19(1): 130.
[13] CHEN Y, LIN Y, SHU Y Q, et al. Interaction between N⁶-methyladenosine (m⁶A) modification and noncoding RNAs in cancer[J]. Mol Cancer, 2020, 19(1): 94.
[14] LEE Y J, CHOE J, PARK O H, et al. Molecular mechanisms driving mRNA degradation by m⁶A modification[J]. Trends Genet, 2020, 36(3): 177-188.
[15] WANG T Y, KONG S, TAO M, et al. The potential role of RNA N⁶-methyladenosine in Cancer progression[J]. Mol Cancer, 2020, 19(1): 88.
[16] ZHANG Z, WANG M, XIE W, et al. m⁶A modification and its regulatory proteins in thyroid cancer: a comprehensive analysis[J].Front Endocrinol, 2021, 12: 721331.
[17] SONG R H, ZHAO J, GAO C Q, et al. Inclusion of ALKBH5 as a candidate gene for the susceptibility of autoimmune thyroid disease[J]. Adv Med Sci, 2021, 66(2): 351-358.
[18] LI W, HUANG G, WEI J R, et al. ALKBH5 inhibits thyroid cancer progression by promoting ferroptosis through TIAM1-Nrf2/HO-1 axis[J]. Mol Cell Biochem, 2023, 478(4): 729-741.
[19] JI X Y, LV C Z, HUANG J P, et al. ALKBH5-induced circular RNA NRIP1 promotes glycolysis in thyroid cancer cells by targeting PKM2[J]. Cancer Sci, 2023, 114(6): 2318-2334.
[20] LIU H, LYU H, JIANG G M, et al. ALKBH5-Mediated m⁶A demethylation of glut4 mRNA promotes glycolysis and resistance to HER²-targeted therapy in breast cancer[J]. Cancer Res, 2022, 82(21): 3974-3986.
[21] ZHAI J N, CHEN H R, WONG C C, et al. ALKBH5 drives immune suppression via targeting AXIN2 to promote colorectal cancer and is a target for boosting immunotherapy[J]. Gastroenterology, 2023, 165(2): 445-462.
[22] GALLUZZI L, VITALE I, AARONSON S A, et al. Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018[J]. Cell Death Differ, 2018, 25(3): 486-541.
[23] KASTENHUBER E R, LOWE S W. Putting P53 in context[J]. Cell, 2017, 170(6): 1062-1078.

Intracellular and cellular functions of ALKBH5 in thyroid cancer cells

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