Next, we utilized a HBx-expressing metastatic HCC cell line, MHCC97-H, which showed lung metastasis, to measure the impact of miR-148a on metastasis. The quantity of the intrahepatic nodules and nodules spread through the entire pulmonary region was clearly decreased in the miR-148a¨Cexpressing group in contrast with that in empty vector group . In the 3-dimensional optimum intensity projection and PET photographs, lung-to-blood or liver-to-blood radioactivity in the miR-148a¨C expressing group was significantly decrease than that in manage group. Histologic evaluation to the lungs and livers confirmed the metastasis foci . The amount of tumor foci observed inside the lungs or livers within the miR-148a group was considerably decrease than that within the empty vector group . These findings strongly supported the function of miR-148a like a suppressor of tumor dissemination.
HPIP increases hepatoma cell proliferation, migration, and invasion and promotes EMT by way of regulation of mTOR signaling. Considering the fact that miR- 148a exerts its function as a result of inhibition of HPIP, we determined whether HPIP has opposite functions of miR-148a from the regulation of HCC cell proliferation, migration, and invasion too as EMT. As anticipated, HPIP overexpression c-Raf inhibitor in HepG2 cells promoted cell proliferation , accompanied by elevated amounts of phosphorylation of mTOR, S6K1, and 4E-BP1 and improved expression of c-myc and cyclin D1 . On the other hand, treatment method with the mTOR inhibitor rapamycin abolished the capacity of HPIP to manage cell proliferation too as the mTOR pathway molecules . A related trend was obtained in migration and invasion assays .
Contrary to results found with miR-148a, HPIP enhanced EMT, with enhanced expression of N-cadherin, Vimentin, and Snail and decreased expression of E-cadherin . The observed EMT results could possibly be reversed by rapamycin, suggesting that HPIP promotes EMT by means of regulation of mTOR signaling. Also, HPIP knockdown had equivalent results selleckchem this article to miR-148a overexpression about the regulation of hepatoma cell proliferation, invasion, and EMT and abolished the means of miR-148a to manage these effects . The knockdown results could be rescued by siRNA-resistant HPIP expression. These information indicate that HPIP may be a important mediator of miR-148a function. Also, AKT and ERK1/2 were demanded for miR-148a/HPIP modulation of EMT because inhibition of AKT and ERK1/2 abolished the potential of miR-148a/HPIP to regulate EMT .
Expression of miR-148a and HPIP and correlation among miR-148a, HPIP, and HBV infection in human HCC samples. First, we assessed the miR-148a expression amounts in a HCC cohort consisting of 52 pairs of primary HCC and their corresponding nontumorous livers by real-time RT-PCR. Compared with their corresponding nontumorous counterparts, miR-148a expression was reduced in liver cancer tissues .