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Published on: January 22, 2021
Effect of NS-398 on colon cancer cells
Xiao-Qing Jia1, Ning Zhong, Li-Hui Han
1Department of Gastroenterology, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China. xiaoqingjia@126.com
Aim:
To study the effect of NS-398, a selective cyclooxygenase-2 (COX-2) inhibitor, on invasion of colon cancer cell line HT-29 in vitro and to explore its mechanisms.
Methods:
Invasive behaviors of the malignant colon cancer cell line HT-29 were investigated in this study. Expressions of COX-2 and CD44v6 in HT-29 cells were detected by flow cytometry. Cellular survival rate was determined by MTT assay. The invasive capacity was quantified by a modified Boyden chamber model. Alterations of cytoskeleton component F-actin were observed by confocal laser scanning microscope.
Results:
Flow cytometry analysis showed that COX-2 was highly expressed in HT-29 cells. The invasive capability of HT-29 cells could be greatly inhibited by NS-398 at the experimental concentrations of 0.1, 1.0 and 10 micormol/L with an inhibitory rate of 22.74%, 42.35% and 58.61% (P<0.01), respectively. MTT assay showed that NS-398 at the experimental concentrations had no significant influence on cellular viability, indicating that such anti-invasive effects had no relationship with cytotoxicity. F-actin was mainly distributed around nuclei forming annular structure in HT-29 cells. After exposure to NS-398 of 10 micromol/L, the annular structure around nuclei disappeared and the fluorescence intensity of F-actin decreased obviously. Treatment with NS-398 could down-regulate the expression of CD44v6 as well.
Conclusion:
NS-398 has anti-invasive effects on colon cancer HT-29 cells in vitro, which may be mediated by a novel mechanism of disruption of cytoskeleton. Down-regulation of CD44v6 expression may be related to alterations of cytoskeleton.
Insights
The drug NS-398 effectively inhibits colon cancer cell invasion by disrupting the cytoskeleton and down-regulating CD44v6 expression. This study reveals a novel mechanism for NS-398
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Colon cancer cell invasion is a critical factor in metastasis.
- Cyclooxygenase-2 (COX-2) is implicated in cancer progression.
- Targeting COX-2 offers a potential therapeutic strategy for colon cancer.
Purpose of the Study:
- To investigate the anti-invasive effects of NS-398, a selective COX-2 inhibitor, on HT-29 colon cancer cells.
- To elucidate the underlying mechanisms of NS-398's action on colon cancer cell invasion.
Main Methods:
- HT-29 colon cancer cells were used to assess invasive behaviors in vitro.
- Expression of COX-2 and CD44v6 was analyzed via flow cytometry.
- Cellular viability was determined by MTT assay.
- Invasion was quantified using a modified Boyden chamber model.
- Cytoskeleton alterations (F-actin) were visualized by confocal microscopy.
Main Results:
- NS-398 significantly inhibited HT-29 cell invasion in a dose-dependent manner (22.74% to 58.61% inhibition).
- NS-398 did not affect cellular viability, indicating non-cytotoxic anti-invasive effects.
- NS-398 disrupted the F-actin cytoskeleton structure and reduced its fluorescence intensity.
- NS-398 treatment led to down-regulation of CD44v6 expression.
Conclusions:
- NS-398 exhibits significant anti-invasive effects on HT-29 colon cancer cells in vitro.
- The anti-invasive mechanism may involve disruption of the cytoskeleton.
- Down-regulation of CD44v6 expression is potentially linked to NS-398-induced cytoskeleton alterations.
