Nanomechanica Signature Alteration Induced by Neuropilin-1 in Hepatocellular Carcinoma Cells

Neuropilin-1 (NRP1) is a type I transmembrane glycoprotein that was originally found playing a role in neuronal axon guidance and embryonic angiogenesis. NRP1 can promote various aspects of tumorigenesis, such as angiogenesis, cell survival, migration, invasion and chemo-resistance, but little is known about the nanomechanica signature of NRP1 inregulating hepatocellular carcinoma(HCC) cells. Our study found that NRP1 was overexpressed in HCC and high expression of NRP1 was associated with poor prognosis in HCC patients.but there are no correlation between NRP1 and liver cirrhosis. In vivo and vitro experiments knockdown of NRP1 reduced proliferation and invasion of HCC. We quantified the Young's modulus, RMS residuals of HCC cell lines and normal liver cell line L02 using Atomic force microscopy (AFM). AFM data indicated that Young's modulus decreased in highly metastatic HCC cells, which is consistent with the verification results of NRP1 knockdown. Our analyses revealed that down regulation of NRP1 negatively correlated with increased Young's modulus in cells exhibiting differing degrees of invasiveness. Our molecular data showed that NRP1 activated AKT, ERK1/2, and epithelial-mesenchymal transition (EMT)-related signals in HCC cells and promoted tumor apoptosis and invasion. As an important biophysical property of cells, the membrane stiffness of HCC cells was further quantified by AFM force measurements, which indicated that these cells became much stiffer after NRP1 knockdown. These results collectively illuminated that Neuropilin-1 alterrated namomechanica signature in hepatocellular carcinoma cells and can be a potential therapeutic agent for HCC. Funding Statement: The present study was supported by the National Natural Science Fund of China (grant nos. 31570509 and 81702326), Science and Technology Major Project of Gansu province(1602FKDA001), the Talent innovation and Entrepreneurship Program of Lanzhou City (grant no. 2016-RC57) and the science and technology project of Chenguan District (2017SHFZ0014); Declaration of Interests: The authors declare no conflicts of interest. Ethics Approval Statement: The study was approved by the Zhongshan Hospital Ethics Committee and informed consent was obtained from each patient under Institutional Reviewer Board protocols. The study protocol conforms to the ethical guidelines of the 1975 Declaration of Helsinki as reflected in a prior approval by the institution's human research committee.