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Real-Time Cell Analysis Reveals Distinct Roles of S100A4 in Regulating Proliferation, Migration, and Invasion of JAR Choriocarcinoma Cells

Yu, B.; Ding, H.-G.; Zhang, F.; Lin, H.-M.; Xia, G.-Y.; Jiang, Y.-J.; Zhao, J.; Li, G.-P.; Ding, J.-L.; Ding, N.; Zhang, X.-Y.; Pan, H.-T.; Ying, P.; He, Y.

2026-02-28 cell biology
10.64898/2026.02.26.708094 bioRxiv
Show abstract

S100A4, a metastasis promoting calcium binding protein, drives tumor progression through pleiotropic mechanisms, yet its context dependent functions in gestational malignancies remain elusive. To dynamically decode its role in choriocarcinoma pathogenesis, we leveraged label free real time cell analysis (RTCA) to profile malignant phenotypes in JAR cells following siRNA mediated S100A4 silencing, complemented by apoptosis assessment and targeted signaling profiling. Efficient knockdown (verified by qPCR/Western blotting) significantly attenuated cellular proliferation (96 hr cell index slope decreased vs. scramble control; p<0.01) and suppressed migration capacity (p<0.01). Critically, S100A4 depletion did not induce apoptosis (flow cytometry and cleaved caspase 3/9 blotting confirmed no significant change), and invasion through Matrigel coated membranes remained statistically unaltered despite comparable experimental rigor. Mechanistically, S100A4 silencing triggered adaptive signaling rewiring: IRS1 and PI3K expression were elevated, Akt1 was suppressed, while MEK1/2 remained unchanged suggesting compensatory pathway activation.

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