LASS2/TMSG1 gene silencing promotes the invasiveness and metastatic of human prostatic carcinoma cells through increase in vacuolar ATPase activity.
- Author:
Xiaoyan XU
1
;
Jiangfeng YOU
1
;
Fei PEI
2
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis; Cell Cycle; Cell Line, Tumor; Gene Silencing; Humans; Hydrogen-Ion Concentration; Lymphatic Metastasis; Male; Matrix Metalloproteinase 2; metabolism; Matrix Metalloproteinase 9; metabolism; Membrane Proteins; genetics; metabolism; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Neoplasm Transplantation; Prostatic Neoplasms; genetics; metabolism; pathology; RNA, Small Interfering; genetics; Sphingosine N-Acyltransferase; genetics; metabolism; Transfection; Tumor Burden; Tumor Suppressor Proteins; genetics; metabolism; Up-Regulation; Vacuolar Proton-Translocating ATPases; metabolism
- From: Chinese Journal of Pathology 2014;43(3):177-183
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effects of LASS2/TMSG1 silencing on the growth, invasion and metastasis of prostate carcinoma cells and to investigate the related molecular mechanisms.
METHODSLASS2/TMSG1 expression of human prostate carcinoma cell line with low metastatic potentiality (PC-3M-2B4 cells) was knocked down using DNA vector-based small interfering RNA (shRNA), followed by evaluations of tumor cell invasion and metastasis.
RESULTSA stable PC-3M-2B4 cell line with expression of LASS2/TMSG1-shRNA was successfully established. MTT assay showed PC-3M-2B4 cells exhibited a strong proliferation after transfection of LASS2/TMSG1-shRNA.LASS2/TMSG1-shRNA transfected clones demonstrated an increased clonogenicity by soft agar colony formation assay and a significant increase of tumor cell invasion by matrigel invasion study.Flow cytometry showed that after LASS2/TMSG1 gene silencing, the apoptotic rate of PC-3M-2B4 cell significantly decreased (P<0.01) without significant cell cycle change (P>0.05).Eight weeks after implantation into subcutaneous tissues in BAL B/c (nu+) mice, the size and weight of sh-LASS2/TMSG1 xenografts were significantly larger than those of the control group (P<0.05).Nuclear proliferation index of the subcutaneous tumor was also higher in the LASS2/TMSG1 shRNA group than those in the control group. Lymph node metastasis was observed in 5 of 6 mice of LASS2/TMSG1 shRNA group and only 1 of 6 of the control group. V-ATPase activity, activities of secreted MMP-2 and MMP-9 and extracellular hydrogen ion concentration were significantly increased in LASS2/TMSG1-shRNA group compared with the control group (P<0.05).
CONCLUSIONSilencing of LASS2/TMSG1 promotes the growth, invasion and metastasis of prostate cancer cells through up-regulation of V-ATPase activity, indicating that LASS2/TMSG1 is a tumor metastasis suppressor gene.
