Knockdown of Mgat5 inhibits CD133+ human pulmonary adenocarcinoma cell growth in vitro and in vivo.

Abstract

PURPOSE:

In spite of many therapeutic advances, the prognosis of lung cancer remains poor. Therefore, understanding the molecular mechanisms underlying cancer progression, invasion and metastasis is needed. Accumulating evidence indicate that N-acetylglucosaminyltransferase V (Mgat5 or GnT-V) is involved in cancer development. The purpose of this study was to characterize the expression and function of Mgat5 in CD133+ pulmonary adenocarcinoma cells.

METHODS:

CD133+ pulmonary adenocarcinoma cells were separated by magnetic activated cell sorting (MACS) from excised pulmonary adenocarcinoma specimens from 10 patients. Expression of Mgat5 in CD133+ cells was detected by fluorescent quantitative RT-PCR (FQRT-PCR) and Western blot. Subsequently, CD133+ cells were transfected with specific siRNA of Mgat5 to evaluate the effects of Mgat5 inhibition on cancer cell growth in vivo and in vitro.

RESULTS:

Expression of Mgat5 was 1.2-fold and 1.4-fold higher in CD133+cells than in CD133- cells detected by FQRT-PCR and Western Blot, respectively (p < 0.05). The L-PHA binding assay also showed higher reactivity in CD133+ cells than in CD133- cells. In addition, Mgat5-specific siRNA efficiently knocked down the expression of Mgat5 in CD133+ cells. Interestingly, downregulation of Mgat5 resulted in significant inhibition of cancer cell growth in vitro and in vivo.

CONCLUSION:

Mgat5 is expressed at a relatively high level in CD133+ lung adenocarcinoma cells, and knockdown of Mgat5 in CD133+ cells inhibits cancer cell growth both in vitro and in vivo. These findings suggest Mgat5 may play an important role during oncogenesis, identifying a potential therapeutic target for pulmonary adenocarcinoma.

• Authors: Xuefeng Zhou, Haidan Chen, Qilong Wang, Li Zhang, Jinping Zhao
• Journal: Clinical and investigative medicine. Medecine clinique et experimentale (Clin Invest Med) Vol. 34 Issue 3 Pg. E155-62 (Jun 01 2011) ISSN: 1488-2353 [Electronic] Canada
• PMID: 21631992 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• AC133 Antigen
• Antigens, CD
• Glycoproteins
• PROM1 protein, human
• Peptides
• Prom1 protein, mouse
• RNA, Small Interfering
• N-Acetylglucosaminyltransferases
• alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase

Topics

• AC133 Antigen
• Adenocarcinoma (genetics, metabolism, therapy)
• Adenocarcinoma of Lung
• Animals
• Antigens, CD (metabolism)
• Blotting, Western
• Cell Proliferation
• Female
• Glycoproteins (metabolism)
• Humans
• Lung Neoplasms (genetics, metabolism, therapy)
• Male
• Mice
• Mice, SCID
• N-Acetylglucosaminyltransferases (genetics, metabolism)
• Peptides (metabolism)
• RNA, Small Interfering
• Reverse Transcriptase Polymerase Chain Reaction
• Tumor Cells, Cultured
• Xenograft Model Antitumor Assays