着丝粒蛋白T在乳腺癌中表达及其启动子区变异的研究

doi:10.3969/j.issn.1004-616x.2022.05.001

Abstract

Abstract: OBJECTIVE: To explore expressions and biological functions of CENP-T in breast cancer,and to analyze the mutation and methylation status in gene promoter sequences.METHODS: Expressions of the CENP-T protein in 26 pairs of breast cancer tissues and adjacent normal breast tissues were detected by immunohistochemical method.Tissue genomic DNA was extracted and sequenced to analyze mutation of CENP-T in the promoter region and methylation levels of CpG islands in the promoter region after bisulfite modification.Western blot was used to detect CENP-T protein expression in three cell lines:MCF10A,MCF7 and MDA-MB-231.The MCF7 cell line was constructed by CENP-T siRNA transient transfection,and effects of the interference were evaluated using Western blot.After the knockdown,expressions of CENP-T,and proliferation ability were examined using the MTT and the plate colony formation assays.Cell cycle distribution and apoptosis were testified by flow cytometry.RESULTS: CENP-T protein expressions in the breast cancer group was significantly lower than that in the adjacent normal breast group (P<0.01).Three mutations of C1417T,C1545T and C1628G were detected in the CENT-P promoter region of breast cancer tissues,and the methylation levels were increased by 49.5%.After CENP-T knockdown,proliferation activities of the MCF7 cells were enhanced (P<0.05),colony formation abilities were enhanced (P<0.01),proportions of cells in the G2/M phase were significantly increased (P<0.01),and apoptosis rates were decreased (P<0.01).CONCLUSION: The down-regulation of CENP-T protein levels promoted proliferations of breast cancer cells which were associated with promoter region mutations and increased methylations.These results indicate that the downregulation of CENP-T expression was related to breast cancer risk.

Key words: CENP-T, breast carcinoma, promoter region, proliferation

CLC Number:

R73-3

BI Jiaxin, WANG Xiaoxue, ZHAO Ruohan, HAN Xu, SONG Jie. Down-expression of CENP-T and variation of its promoter region in BRCA[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(5): 327-334.

References

[1] MUSACCHIO A, DESAI A. A molecular view of kinetochore assembly and function[J]. Biology, 2017, 6(1):5.
[2] LEE M, CHANG J, CHANG S, et al. Asymmetric spindle pole formation in CPAP-depleted mitotic cells[J]. Biochem Biophys Res Commun, 2014, 444(4):644-650.
[3] KIM S, YU H T. Multiple assembly mechanisms anchor the KMN spindle checkpoint platform at human mitotic kinetochores[J]. J Cell Biol, 2015, 208(2):181-196.
[4] SUZUKI A, BADGER B L, WAN X H, et al. The architecture of CCAN proteins creates a structural integrity to resist spindle forces and achieve proper intrakinetochore stretch[J]. Dev Cell, 2014, 30(6):717-730.
[5] FRITZLER M J, KINSELLA T D, GARBUTT E. The CREST syndrome:a distinct serologic entity with anticentromere antibodies[J]. Am J Med, 1980, 69(4):520-526.
[6] HELLWIG D, MÜNCH S, ORTHAUS S, et al. Live-cell imaging reveals sustained centromere binding of CENP-T via CENP-A and CENP-B[J]. J Biophotonics, 2008, 1(3):245-254.
[7] WATANABE R, HIRANO Y, HARA M, et al. Mobility of kinetochore proteins measured by FRAP analysis in living cells[J]. Chromosome Res, 2022, 30(1):43-57.
[8] DONG Q, LIU X L, WANG X H, et al. Ccp1-Ndc80 switch at the N terminus of CENP-T regulates kinetochore assembly[J]. PNAS, 2021, 118(48):e2104459118.
[9] KUKREJA A A, KAVURI S, JOGLEKAR A P. Microtubule attachment and centromeric tension shape the protein architecture of the human kinetochore[J]. Curr Biol, 2020, 30(24):4869-4881.
[10] CORTES-SILVA N, ULMER J, KIUCHI T, et al. CenH3-independent kinetochore assembly in Lepidoptera requires CCAN, including CENP-T[J]. Curr Biol, 2020, 30(4):561-572. e10.
[11] LERA R F, NORMAN R X, DUMONT M, et al. Plk1 protects kinetochore-centromere architecture against microtubule pulling forces[J]. EMBO Rep, 2019, 20(10):e48711.
[12] DING M R, JIANG J Y, YANG F R, et al. Holliday junction recognition protein interacts with and specifies the centromeric assembly of CENP-T[J]. J Biol Chem, 2019, 294(3):968-980.
[13] THAKUR J, HENIKOFF S. CENPT bridges adjacent CENPA nucleosomes on young human α-satellite dimers[J]. Genome Res, 2016, 26(9):1178-1187.
[14] RAGO F, GASCOIGNE K E, CHEESEMAN I M. Distinct organization and regulation of the outer kinetochore KMN network downstream of CENP-C and CENP-T[J]. Curr Biol, 2015, 25(5):671-677.
[15] HORI T, AMANO M, SUZUKI A, et al. CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore[J]. Cell, 2008, 135(6):1039-1052.
[16] SAITO A, MURO Y, SUGIURA K, et al. Low prevalence of autoantibodies to CENP-H,-I,-K,-L,-M,-N,-T and-U in a Japanese cohort of anti-centromere positive samples[J]. Immunopharmacol Immunotoxicol, 2013, 35(1):57-63.
[17] WANG Y, LI J, DONG F, et al. CENP-T regulates both the G2/M transition and anaphase entry by acting through CDH1 in meiotic oocytes[J]. J Cell Sci, 2020, 133(3):jcs238105.
[18] MILAGRE I, PEREIRA C, OLIVEIRA R A, et al. Reprogramming of human cells to pluripotency induces CENPa chromatin depletion[J]. Open Biol, 2020, 10(10):200227.
[19] ATHWAL R K, WALKIEWICZ M P, BAEK S, et al. CENP-a nucleosomes localize to transcription factor hotspots and subtelomeric sites in human cancer cells[J]. Epigenetics Chromatin, 2015, 8:2.
[20] ARIMURA Y, SHIRAYAMA K, HORIKOSHI N, et al. Crystal structure and stable property of the cancer-associated heterotypic nucleosome containing CENP-A and H3.3[J]. Sci Rep, 2014, 4:7115.
[21] WENG M Y, LI L, HONG S J, et al. Clinical significance of CENP-H expression in uterine cervical cancer[J]. Cancer Biol Med, 2012, 9(3):192-196.
[22] HE W L, LI Y H, YANG D J, et al. Combined evaluation of centromere protein H and Ki-67 as prognostic biomarker for patients with gastric carcinoma[J]. Eur J Surg Oncol EJSO, 2013, 39(2):141-149.
[23] 廖长春,范阳华,肖兵,等.缺氧诱导因子-1α与着丝粒蛋白W在人胶质瘤组织及细胞中表达的相关性[J].肿瘤, 2014, 34(11):1045-1051.
[24] 肖峰,颜红柱,邹珏,等.子宫颈鳞状上皮内病变和鳞状细胞癌中CENP-F、Ki-67的表达及意义[J].临床与实验病理学杂志, 2017, 33(5):562-564.
[25] CHEN Y, XU R Y, RUZE R, et al. Construction of a prognostic model with histone modification-related genes and identification of potential drugs in pancreatic cancer[J]. Cancer Cell Int, 2021, 21(1):291.

Down-expression of CENP-T and variation of its promoter region in BRCA

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