ITSN1-S SH3 domains regulate human glioblastoma U87 cell pro-liferation / 中国肿瘤临床

ABSTRACT

Objective:To investigate the functions of the ITSN1-S SH3 domains in U87 glioblastoma cell proliferation and to de-termine the underlying molecular mechanism. Methods: A recombinant lentiviral vector with an mGFP label was constructed. EH1-EH2, EH1-EH2-CC, and ITSN1-S genes were amplified using polymerase chain reaction and then cloned into recombinant lenti-viral vectors. The four lentiviral plasmids were packaged using HEK 293T cells and subsequently used to infect U87 cells. Stable cells were screened using puromycin and separately labeled as vector/U87, EH1-EH2/U87, EH1-EH2-CC/U87, and ITSN1-S/U87. Western blotting was used to detect the expression of each protein. Proliferation and soft agar assays were performed to detect cell proliferation. Results:In the proliferation and soft agar assays, the proliferation capacity of the ITSN1-S/U87 cells was clearly enhanced compared with those of the vector/U87, EH1-EH2/U87, and EH1-EH2-CC/U87 cells (P0.05). On the 6th day, the vector/U87, EH1-EH2/U87, EH1-EH2-CC/U87, and ITSN1-S/U87 cell numbers were (29.16 ± 1.19) × 104, (22.82 ± 0.94) × 104, (22.17 ± 0.90) × 104, and (21.93 ± 1.15) × 104, respectively. On the 21st day, the number of colony formation in vector/U87, EH1-EH2/U87, EH1-EH2-CC/U87, and ITSN1-S/U87 was (6.37±0.41)×103, (2.65±0.34)×103, (2.23±0.31)×103, and (2.1±0.29)×103, respectively . Conclusion:ITSN1-S overexpression significantly promotes U87 cell proliferation. Specifically, the SH3 domains possibly serve vital functions in glioma cell proliferation.