Role And Mechanism Of MiR-605/psmd10 In Intrahepaticcholangiocarcinoma Cell Progression

Background and aimA few studies shows that PSMD10 is involved in the tumorigenicity and metastasis of some malignancies, and it is a new kind of proto-oncogene. Prvious study demonstrated that PSMD10 is highly expressed in cholangiocarcinoma tissue and cell lines, and closely related to differentiation, TNM stage and metastasis of cholangiocarcinoma. These suggest that PSMD10 gene may play a important role in the growth and metastasis of cholangiocarcinoma, which has the potential in molecular therapy.Materials and methods1, PSMD10 sh RNA expression plasmid were constructed, then transfected into RBE and QBC939 cell lines. Transfection efficiency was detected by semi-quantitative PCR and Western blot; cell proliferation were detemined by MTT and soft AGAR plate clone formation test; cell invasion ability was evaluated by transwell cell chamber experiment; And cells apoptosis were determined.2, mi RNAs, which may targeted to PSMD10, were screened by bioinformatics software. Then mi RNAs overexpression plasmid were constructed and tranfected into RBE cell lines. PSMD10 expression levels were detected by Western blot; mi RNAs expression levels were screened in cancer tissue and normal tissue; Finally, verification of mi RNAs targeted to PSMD10 were performed using dual luciferase report system.3, mi RNA overexpression plasmid was transfected into QBC939 and RBE cells, cell proliferation and invasion ability were evaluated by MTT, soft AGAR cloning formation, Transwell cell chamber experiment, respectively. In animals model, Tumorigenic experiment was made to confirm whether mi RNA could inhibit the growth and metastasis of cholangiocarcinoma.4, PSMD10 was over-expressed in QBC939 and RBE cells transfected with mi RNA, whether the inhibitory effect of the m RNA depends on PSMD10 expression was validated by soft AGAR cloning formation and transwell cell chamber experimen. Overexpression of p53 in RBE cells, at the same time, mi RNA was transfected REB cells with high expression level of p53, the levels of PSMD10 and miRNA were detected through western blot and quantitative PCR.Results1. silence of PSMD10 could promote cholangiocarcinoma cell apoptosis, inhibit the proliferation and invasion of cholangiocarcinoma cells;2. mi R- 559, mi R- 605, mi R- 875-3 p and mi R – 1254 were found by bioinformatic screen, which may target to PSMD10; in them, miR- 605 and mi R- 875-3p could significantly decrease the expression levels of PSMD10; Verification results in clinical samples show that miR- 605 was negative correlate with PSMD10; Dual luciferase report experiments confirmed it.3. In vitro, overexpression of miR- 605 could promote cell apoptosis, inhibit the proliferation and invasion of cholangiocarcinoma; In vivo, overexpression of mi R-605 significantly inhibits the subcutaneous growing tumor growth.4. Inhibition of mi R-605 on cholangiocarcinoma depends on PSMD10; PSMD10 is involved in P53- mi R- 605 signal network.Conclusionsmi R-605, which target to PSMD10 in intrahepatic cholangiocarcinoma, play an important role in the growth and metastasis of cells, and P53-miR-605- PSMD10 signal network may be involved in ICC progression.