Knockdown of SASS6 reduces growth of MDA‑MB‑231 triple‑negative breast cancer cells through arrest of the cell cycle at the G2/M phase

Du,Lili; Jing,Jiexian; Wang,Yan; Xu,Xiaoqin; Sun,Ting; Shi,Yanchun; Wang,Weigang; Tian,Baoguo; Han,Cunzhi; Zhao,Xianwen; Chang,Huibo

doi:10.3892/or.2021.8052

Knockdown of SASS6 reduces growth of MDA‑MB‑231 triple‑negative breast cancer cells through arrest of the cell cycle at the G2/M phase

Authors:
- Lili Du
- Jiexian Jing
- Yan Wang
- Xiaoqin Xu
- Ting Sun
- Yanchun Shi
- Weigang Wang
- Baoguo Tian
- Cunzhi Han
- Xianwen Zhao
- Huibo Chang
View Affiliations

Affiliations: Department of Etiology, Shanxi Cancer Hospital, Taiyuan, Shanxi 030013, P.R. China, Department of Biochemistry and Immunology, Capital Institute of Pediatrics, Beijing 100020, P.R. China
Published online on: April 13, 2021 https://doi.org/10.3892/or.2021.8052
Article Number: 101

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Spindle assembly abnormal protein 6 homolog (SASS6) is crucial for centriole duplication; however, the role of SASS6 in the proliferation of cancer cells remains unclear. In the present study, the expression and functional role of SASS6 in triple negative breast cancer (TNBC) was assessed. Immunohistochemical staining was performed using an anti‑SASS6 antibody in TNBC and normal tissues. Lentivirus‑mediated RNA interference was used to knockdown SASS6 in MDA‑MB‑231 TNBC cells. Cell viability was determined using an MTT assay, and cell cycle distribution and apoptosis were measured using flow cytometry. Additionally, PathScan intracellular signaling arrays were used to detect the presence of intracellular signaling molecules. The results revealed that SASS6 expression was increased in TNBC tissues compared with the control tissue. Moreover, SASS6 knockdown significantly suppressed the growth of MDA‑MB‑231 cells. MDA‑MB‑231 cell cycle progression was arrested at the G2/M phase and cyclin dependent kinase 1 (CDK1), cyclin B1 and PCNA expression in MDA‑MB‑231 cells was decreased following SASS6 knockdown. Furthermore, the phosphorylation of STAT3, BAD and rpS6 was reduced following SASS6 knockdown. A strong correlation between SASS6 and CDK1 expression was observed in TNBC tissues based on immunohistochemical staining analysis (R=0.989; P<0.001). In conclusion, the present study revealed the crucial role of SASS6 in promoting MDA‑MB‑231 cell growth, regulating cell cycle progression and its ability to downregulate the CDK1/cyclin B1 signaling pathway, thus highlighting the potential of SASS6 as a therapeutic target for treatment of TNBC, and merits further investigation in animal models or in preclinical and clinical studies.

View Figures

View References

1	Sharma P: Update on the treatment of early-stage triple-negative breast cancer. Curr Treat Options Oncol. 19:222018. View Article : Google Scholar : PubMed/NCBI
2	De Laurentiis M, Cianniello D, Caputo R, Stanzione B, Arpino G, Cinieri S, Lorusso V and De Placido S: Treatment of triple negative breast cancer (TNBC): Current options and future perspectives. Cancer Treat Rev. 36 (Suppl 3):S80–S86. 2010. View Article : Google Scholar
3	Rivera-Rivera Y and Saavedra HI: Centrosome-a promising anti-cancer target. Biologics. 10:167–176. 2016.PubMed/NCBI
4	Raff JW: Centrosomes and cancer: Lessons from a TACC. Trends Cell Biol. 12:222–225. 2002. View Article : Google Scholar : PubMed/NCBI
5	Salisbury JL: The contribution of epigenetic changes to abnormal centrosomes and genomic instability in breast cancer. J Mammary Gland Biol Neoplasia. 6:203–212. 2001. View Article : Google Scholar : PubMed/NCBI
6	Korzeniewski N, Hohenfellner M and Duensing S: The centrosome as potential target for cancer therapy and prevention. Expert Opin Ther Targets. 17:43–52. 2013. View Article : Google Scholar : PubMed/NCBI
7	Chan JY: A clinical overview of centrosome amplification in human cancers. Int J Biol Sci. 7:1122–1144. 2011. View Article : Google Scholar : PubMed/NCBI
8	Pihan GA, Purohit A, Wallace J, Knecht H, Woda B, Quesenberry P and Doxsey SJ: Centrosome defects and genetic instability in malignant tumors. Cancer Res. 58:3974–3985. 1998.PubMed/NCBI
9	Leidel S, Delattre M, Cerutti L, Baumer K and Gönczy P: SAS-6 defines a protein family required for centrosome duplication in C. elegans and in human cells. Nat Cell Biol. 7:115–125. 2005. View Article : Google Scholar : PubMed/NCBI
10	Yoshiba S, Tsuchiya Y, Ohta M, Gupta A, Shiratsuchi G, Nozaki Y, Ashikawa T, Fujiwara T, Natsume T, Kanemaki MT and Kitagawa D: HsSAS-6-dependent cartwheel assembly ensures stabilization of centriole intermediates. J Cell Sci. 132:jcs2175212019. View Article : Google Scholar : PubMed/NCBI
11	Strnad P, Leidel S, Vinogradova T, Euteneuer U, Khodjakov A and Gönczy P: Regulated HsSAS-6 levels ensure formation of a single procentriole per centriole during the centrosome duplication cycle. Dev Cell. 13:203–213. 2007. View Article : Google Scholar : PubMed/NCBI
12	Shinmura K, Kato H, Kawanishi Y, Nagura K, Kamo T, Okubo Y, Inoue Y, Kurabe N, Du C, Iwaizumi M, et al: SASS6 overexpression is associated with mitotic chromosomal abnormalities and a poor prognosis in patients with colorectal cancer. Oncol Rep. 34:727–38. 2015. View Article : Google Scholar : PubMed/NCBI
13	Annika W, Anja MB, Kereshmeh T, Justus PB, Paul DD, Michael S, Raymund EH, Matthias WB, Pamela LS and Reiner S: Selective isolation and characterization of primary cells from normal breast and tumors reveal plasticity of adipose derived stem cells. Breast Cancer Res. 18:322016. View Article : Google Scholar
14	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
15	Borsotti C, Borroni E and Follenzi A: Lentiviral vector interactions with the host cell. Curr Opin Virol. 21:102–108. 2016. View Article : Google Scholar : PubMed/NCBI
16	Zha J, Harada H, Osipov K, Jockel J, Waksman G and Korsmeyer SJ: BH3 domain of BAD is required for heterodimerization with BCL-XL and pro-apoptotic activity. J Biol Chem. 26(272): 24101–24104. 1997. View Article : Google Scholar
17	Comartin D, Gupta GD, Fussner E, Coyaud É, hasegan M, Archinti M, Cheung SW, Pinchev D, Lawo S, Raught B, et al: CEP120 and SPICE1 cooperate with CPAP in centriole elongation. Curr Biol. 23:1360–1366. 2013. View Article : Google Scholar : PubMed/NCBI
18	Xu X, Huang S, Zhang B, Huang F, Chi W, Fu J, Wang G, Li S, Jiang Q and Zhang C: DNA replication licensing factor Cdc6 and Plk4 kinase antagonistically regulate centrosome duplication via Sas-6. Nat Commun. 8:151642017. View Article : Google Scholar : PubMed/NCBI
19	Arquint C, Sonnen KF, Stierhof YD and Nigg EA: Cell-cycle-regulated expression of STIL controls centriole number in human cells. J Cell Sci. 125:1342–1352. 2012. View Article : Google Scholar : PubMed/NCBI
20	Lin YC, Chang CW, Hsu WB, Tang CJ, Lin YN, Chou EJ, Wu CT and Tang TK: Human microcephaly protein CEP135 binds to hSAS-6 and CPAP, and is required for centriole assembly. EMOJ. 32:1141–1154. 2013.
21	Puklowski A, Homsi Y, Keller D, May M, Chauhan S, Kossatz U, Grünwald V, Kubicka S, Pich A, Manns MP, et al: The SCF-FBXW5 E3-ubiquitin ligase is regulated by PLK4 and targets HsSAS-6 to control centrosome duplication. Nat Cell Biol. 13:1004–1009. 2011. View Article : Google Scholar : PubMed/NCBI
22	Keller D, Orpinell M, Olivier N, Wachsmuth M, Mahen R, Wyss R, Hachet V, Ellenberg J, Manley S and Gönczy P: Mechanisms of HsSAS-6 assembly promoing centriole formation in human cells. J Cell Biol. 204:697–712. 2014. View Article : Google Scholar : PubMed/NCBI
23	Johnson A and O'Donnell M: Cellular DNA replicases: Components and dynamics at the replication fork. Annu. Rev. Biochem. 74:283–315. 2005.
24	Bromberg JF, Wrzeszcznska MH, Devgan G, Zhao Y, Pestell RG, Albanese C and Darnell JE Jr: Stat3 as an oncogene. Cell. 98:295–303. 1999. View Article : Google Scholar : PubMed/NCBI
25	William MB, Jin XH, Lin HJ, Huang M, Liu R, Reynolds RK and Lin J: Inhibition of constitutively active Stat3 suppresses growth of human ovarian and breast cancer cells. Oncogene. 20:7925–7934. 2001. View Article : Google Scholar
26	Catlett-Falcone R, Landowski T, Oshiro M, Turkson J, Levitzki A, Savino R, Ciliberto G, Moscinski L, Fernández-Luna JL, Nuñez G, et al: Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity. 10:105–115. 1999. View Article : Google Scholar : PubMed/NCBI
27	Shen Y, Devgan G, Darnell JE Jr and Bromberg JF: Constitutively activated Stat3 protects fibroblasts from serum withdrawal and UV induced apoptosis and antagonizes the proapoptotic effects of activated Stat1. Proc Natl Acad Sci USA. 98:1543–1548. 2001. View Article : Google Scholar : PubMed/NCBI
28	Garcia R, Yu CL, Hudnall A, Catlett R, Nelson KL, Smithgall T, Fujita DJ, Ethier SP and Jove R: Constitutive activation of Stat3 in fibroblasts transformed by diverseoncoproteins and in breast carcinoma cells. Cell Growth Differ. 8:1267–1276. 1997.PubMed/NCBI
29	Roshan S, Liu YY, Banafa A, Chen HJ, li KX, Yang GX, He GY and Chen MJ: Fucoidan induces apoptosis of HepG2 cells by down-regulating p-Stat3. J Huazhong Univ Sci Technolog Med Sci. 34:330–336. 2014. View Article : Google Scholar : PubMed/NCBI
30	Roux PP, Shahbazian D, Vu H, Holz MK, Cohen MS, Taunton J, Sonenberg N and Blenis J: RAS/ERK signaling promotes site-specific ribosomal protein S6 phosphorylation via RSK and stimulates cap-dependent translation. J Biol Chem. 282:14056–14064. 2007. View Article : Google Scholar : PubMed/NCBI
31	Meyuhas O: Ribosomal protein S6 phosphorylation: Four decades of research. Int Rev Cell Mol Biol. 320:41–73. 2015. View Article : Google Scholar : PubMed/NCBI
32	Akar U, Ozpolat B, Mehta K, Lopez-Berestein G, Zhang D, Ueno NT, Hortobagyi GN and Arun B: Targeting p70S6K prevented lung metastasis in a breast cancer xenograft model. Mol Cancer Ther. 9:1180–1187. 2010. View Article : Google Scholar : PubMed/NCBI
33	Zha J, Harada H, Yang E, Jockel J and Korsmeyer SJ: Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell. 87:619–628. 1996. View Article : Google Scholar : PubMed/NCBI
34	Yang E, Zha J, Jockel J, Boise LH, Thompson CB and Korsmeyer SJ: Bad, a heterodimeric partner for Bcl-XL and Bcl-2, displaces Bax and promotes cell death. Cell. 80:285–291. 1995. View Article : Google Scholar : PubMed/NCBI
35	Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA, MacGregor GR, Thompson CB and Korsmeyer SJ: Proapoptotic BAX and BAK: A requisite gateway to mitochondrial dysfunction and death. Science. 292:727–730. 2001. View Article : Google Scholar : PubMed/NCBI