Abstract
Purpose
To investigate the effect of shRNA-regulated S100A4 expression on the proliferation and apoptosis in KLE endometrial cancer cells.
Methods
S100A4-OVER and S100A4-shRNA were transfected into KLE endometrial cancer cells using lentiviral sh-RNA technology. Passive OVER-NC cell line and shRNA-NC cell line were used as a negative control group and non-transfected Control cell line as a blank control group. After 48 h of transfection, the expressions of S100A4 and protein were detected by real-time fluorescence quantitative PCR and Western blotting, respectively. CCK-8 detection and flow cytometer were used to detect cell proliferation and apoptosis, respectively.
Results
Compared with the normal control group and the negative control group, the transfection efficiency and shRNA targeting of the shRNA-interfered S100A4 gene were verified at the levels of mRNA and protein expression. The expression of the disrupted S100A4 gene at S100A4 mRNA and protein levels in endometrial cancer cells was determined. The proliferation efficiency of KLE cells in S100A4-OVER group was significantly higher than that in other four groups; the proliferation rate of S100A4-shRNA cells decreased slightly;, the apoptotic rate of KLE cells in S100A4-shRNA group increased significantly, and the apoptotic rate of KLE cells in S100A4-OVER group decreased compared with NC group.
Conclusion
Specific regulation of S100A4 gene expression:, the enhanced expression of the S100A4 gene may promote the proliferation of KLE endometrial cancer cells; the inhibited expression of the S100A4 gene may promote the apoptosis of KLE endometrial cancer cells. S100A4 expression is closely related to the biological characteristics of endometrial cancer.
Similar content being viewed by others
References
Chen H, Xu C, Jin Q, Liu Z. S100 protein family in human cancer. Am J Cancer Res. 2014;4:89–115.
Donato R, Cannon BR, Sorci G, et al. Functions of S100 proteins. Curr Mol Med. 2013;13:24–57.
Ismail NI, Kaur G, Hashim H, Hassan MS. S100A4 over-expression proves to be independent marker for breast cancer progression. Cancer Cell Int. 2008;8:12.
Wang YY, Ye ZY, Zhao ZS, et al. High-level expression of S100A4 correlates with lymph node metastasis and poor prognosis in patients with gastric cancer. Ann Surg Oncol. 2010;17:89–97.
Kwak JM, Lee HJ, Kim SH, et al. Expression of protein S100A4 is a predictor of recurrence in colorectal cancer. World J Gastroenterol. 2010;16:3897–904.
Min HS, Choe G, Kim SW, et al. S100A4 expression is associated with lymph node metastasis in papillary microcarcinoma of the thyroid. Mod Pathol. 2008;21:748–55.
Tsuna M, Kageyama SI, Fukuoka J, et al. Significance of S100A4 as a prognostic marker of lung squamous cell carcinoma. Anticancer Res. 2009;29:2547–54.
Davies BR, O'Donnell M, Durkan GC, et al. Expression of SI00A4 protein is associated with metastasis and reduced survival in human bladder cancer. J Pathol. 2002;196:292–9.
Kikuchi N, Horiuchi A, Osada R, et al. Nuclear expression of S100A4 is associated with aggressive behavior of epithelial ovarian carcinoma: an important autocrine/paracrine factor in tumor progression. Cancer Sci. 2006;97:1061–9.
Chong H, Lee J, Yoon M, et al. Prognostic value of cytoplasmic expression of S100A4 protein in endometrial carcinoma. Oncol Rep. 2014;31:2701–7.
Donato R. S100: a multigenic family of calci-um-modulated proteins of the EF-hand type with intracellular and extracellular unctional roles. Int J Biochem Cell Biol. 2001;33:637–68.
Heizmann CW, Fritz G, Schafer BW. S100 proteins: structure, functions and pathology. Front Biosci. 2002;7:d1356–1368.
Boye K, Maelandsmo GM. S100A4 and metastasis: a small actor playing many roles. Am J Pathol. 2010;176:528–35.
Mylona E, Giannopoulou I, Fasomytakis E, Nomikos A, Magkou C, Bakarakos P, Nakopoulou L. The clinicopathologic and prognostic significance of CD44+/CD24(−/low) and CD44−/CD24+ tumor cells in invasive breast carcinomas. Hum Pathol. 2008;7:1096–102.
Liu J, Fu S, Xu Y, Zheng Z. RNA interference targeting inhibition of S100A4 suppresses cell growth and promotes apoptosis in human laryngeal carcinoma Hep-2 cells. Mol Med Rep. 2014;3:1389–94.
Egeland EV, Boye K, Park D, Synnestvedt M, Sauer T, Oslo Breast Cancer Consortium (OSBREAC), Naume B, Borgen E, Mælandsmo GM. Prognostic significance of S100A4-ex-pression and subcellular localization in early-stage breast cancer. Breast Cancer Res Treat. 2017;162:127–37.
Wang H, Shi J, Luo Y, Liao Q, Niu Y, Zhang F, Shao Z, Ding Y, Zhao L. LIM and SH3 protein 1 induces TGFβ-mediated epithelial-mesenchymal transition in human colorectal cancer by regulating S100A4 expression. Clin Cancer Res. 2014;22:5835–47.
Wang XG, Meng Q, Qi FM, Yang QF. Blocking TGF-13 inhibits breast cancer cell invasiveness via ERK/S100A4 signal. Eur Rev Med Pharmacol Sci. 2014;18:3844–53.
Xu X, Su B, Xie C, Wei S, Zhou Y, Liu H, Dai W, Cheng P, Wang F, Xu X, Guo C. Sonic Hedgehog-Glil signaling path—way regulates the epithelial mesenehymal transition (EMT) by me-diating a new target gene, S100A4, in pancreatic cancer cells. PLoS ONE. 2014;9:e96441.
Küper C, Beck FX, Neohofer W. NFAT5-mediated expression of S100A4 contributes to proliferation and migration of renal carcino-ma cells. Front Physiol. 2014;5:293.
Hua J, Chen D, Fu H, Zhang R, Shen W, Liu S, Sun K, Sun X. Short hairpin RNA·mediated inhibition of S100A4 promotes apoptosis and suppresses proliferation of BGC823 gastric cancer cells in vitro and in vivo. Cancer Lett. 2010;292:41–7.
Liu J, Fu S, Xu Y, Zheng Z. RNA interference targeting inhibition of S100A4 suppresses cell growth and promotes apoptosis in human laryngeal carcinoma Hep 2 cells. Mol Med Rep. 2014;10:1389–94.
Yang XC, Wang X, Luo L, Dong DH, Yu QC, Wang XS, Zhao K. RNA interference suppression of A100A4 reduces the growth and metastatic phenotype of human renal cancer cells via NF-kB-dependent MMP-2 and bcl-2pathway. Eur Rev Med Pharmacol Sci. 2013;17:1669–800.
Ma L, Wu AG, Ji SF, Yang HF. Effect of short hairpin RNA-targeted S100A4 on proliferation of breast cancer MCF-7 cells. Chin PLA Postgrad Med Sch. 2010;31:63–6.
Mahon PC, Baril P, Bhakta V, Chelala C, Caulee K, Harada T, Lemoine NR. S100A4 contributes to the suppres-sion of BNIP3 expression, chemoresistance, and inhibition of apoptosis in pancreatic cancer. Cancer Res. 2007;67:6786–95.
Li Q, Dai C, Xue R, Wang P, Chen L, Han Y, Erben U, Qin Z. S100A4 protects myeloid-derived suppressor cells from intrinsic apoptosis via TLR4-ERK1/2 signaling. Front Immunol. 2018;9:388.
Grigorian M, Lukanidin E. Activator of me-tastasis in cancer cells, Mst1/S100A4 protein binds to tumor suppressor protein p53. Genetika. 2003;39:900–8.
Orre LM, Panizza E, Kaminskyy VO, Vernet E, Gräslund T, Zhivotovsky B, Lehtiö J. S100A4 interacts with p53 in the nucleus and promotes p53 degradation. Oncogene. 2013;32:5531–40.
Funding
No funding was received.
Author information
Authors and Affiliations
Contributions
WR wrote the manuscript, and performed PCR and Western blot. YBC was responsible for CCK-8 detection and Flow cytometry. JLS contributed to the observation indexes analysis. The final version was read and adopted by all the authors. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
The study was approved by the Ethical Committee of General Hospital of Northern Theater Command (Heping Campus) and conducted in accordance with the ethical standards.
Informed consent
Not applicable.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Ren, W., Chi, Y.B. & Sun, J.L. Effect of shRNA-mediated regulation of S100A4 gene expression on proliferation and apoptosis of KLE endometrial cancer cells. Clin Transl Oncol 23, 148–154 (2021). https://doi.org/10.1007/s12094-020-02406-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12094-020-02406-7