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[Inhibition of 6A8 alpha-manosidase expression induces decrease of adhesion to laminin and reduction of lamellipodia
Wei Yue1, Fang-tao Zhao, Geng-xian Shi
1Department of Immunology, Institute of Basic Medcial Sciences, CAMS, PUMC, Beijing 100005, China.
Objective:
To investigate the inhibitory effect of 6A8 alpha-manosidase expression on the adhesiveness of CNE-2L2 cells to laminin and the lamellipodia on cell surface.
Methods:
6A8 alpha-manosidase expression was detected by Western blotting. For assaying the adhesion of cells to laminin, cells were incubated in laminin-coated plate at 37 degrees C for 1 h, the adhered cells were stained with crystal purple dissolved in 0.1 mol/L Sodium Citrate/50% ethanol. Absorbance 540 nm was measured. Adhesion rate (R) was calculated according to formula R = AT/A100 x 100%. Here A100 represents 100% adhesion. lamellipodia on cell surface was observed upon a scanning electron microscopy.
Results:
The adhesion rate of two clones (AS1 and AS2) with inhibition of 6A8 alpha-manosidase expression to laminin was 0.447 +/- 0.096 and 0.533 +/- 0.065 respectively. The adhesion rate of three controls with normal expression of 6A8 alpha-manosidase to laminin was 0.78 +/- 0.035, 0.7 +/- 0.05 and 0.80 +/- 0.04 respectively. The difference was significant (P < 0.01). CNE-2L2 cells with normal expression of 6A8 alpha-manosidase was rich in lamellipodia on their surface. Lamellipodia nearly disappeared on the cells with inhibition of 6A8 alpha-manosidase expression.
Conclusions:
Inhibition of 6A8 alpha-manosidase expression results in decrease of adhesion to laminin and reduction of lamellipodia of human nasopharyngeal carcinoma cell CNE-2L2.
Insights
Inhibiting 6A8 alpha-manosidase in nasopharyngeal carcinoma cells significantly reduces their adhesion to laminin and cell surface lamellipodia. This finding impacts understanding of cancer cell invasiveness.
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Context:
- Nasopharyngeal carcinoma (NPC) is a significant health concern.
- Cell adhesion and morphology are critical in cancer metastasis.
- Laminin is a key component of the extracellular matrix involved in cell adhesion.
Purpose:
- To investigate the role of 6A8 alpha-manosidase in regulating CNE-2L2 cell adhesion to laminin.
- To determine the effect of 6A8 alpha-manosidase expression on cell surface lamellipodia formation.
Summary:
- Western blotting confirmed reduced 6A8 alpha-manosidase expression in AS1 and AS2 clones.
- Adhesion assays showed significantly lower cell adhesion rates to laminin in cells with inhibited 6A8 alpha-manosidase (0.447-0.533) compared to controls (0.7-0.8).
- Scanning electron microscopy revealed a near absence of lamellipodia on cells with inhibited 6A8 alpha-manosidase expression, unlike controls.
Impact:
- Inhibition of 6A8 alpha-manosidase decreases the adhesive capacity of CNE-2L2 cells to laminin.
- Reduced 6A8 alpha-manosidase expression leads to a significant reduction in cell surface lamellipodia.
- These findings suggest 6A8 alpha-manosidase as a potential target for modulating NPC cell adhesion and invasiveness.
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