FOLLOWUS
In Vitro antimetastatic effect of Changweiqing (肠胃清) through antiinvasion of hypoxic colorectal carcinoma LoVo cells
- Chinese Journal of Integrative Medicine Vol. 17, Issue 7, Pages: 517-524(2011)
- Affiliations：
Department of Oncology, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai,China
- Author bio：
- Funds：Supported by the National Natural Science Foundation of China (No. 30672683)
DOI：10.1007/s11655-011-0785-0
CLC：Published：2011，
Published Online：3 July 2011，
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Li, J., Fan, Zz., Sun, J. et al. , In Vitro antimetastatic effect of Changweiqing (肠胃清) through antiinvasion of hypoxic colorectal carcinoma LoVo cells., Chin. J. Integr. Med. 17, 517–524 (2011). https://doi.org/10.1007/s11655-011-0785-0
Jing Li, Zhong-ze Fan, Jue Sun, et al. In Vitro antimetastatic effect of Changweiqing (肠胃清) through antiinvasion of hypoxic colorectal carcinoma LoVo cells. [J]. Chinese Journal of Integrative Medicine 17(7):517-524(2011)
Li, J., Fan, Zz., Sun, J. et al. , In Vitro antimetastatic effect of Changweiqing (肠胃清) through antiinvasion of hypoxic colorectal carcinoma LoVo cells., Chin. J. Integr. Med. 17, 517–524 (2011). https://doi.org/10.1007/s11655-011-0785-0 DOI：
Jing Li, Zhong-ze Fan, Jue Sun, et al. In Vitro antimetastatic effect of Changweiqing (肠胃清) through antiinvasion of hypoxic colorectal carcinoma LoVo cells. [J]. Chinese Journal of Integrative Medicine 17(7):517-524(2011) DOI： 10.1007/s11655-011-0785-0.
摘要
To investigate the in vitro effects and the primary mechanisms of Changweiqing (肠胃清
CWQ) on antimetastasis and antiinvasion of hypoxic colon carcinoma cells. In addition
to provide experimental evidence for the Chinese medicinal theory of “strengthening the body’s resistance to eliminate pathogenic factors” in the treatment of colorectal cancer
including its invasion and metastasis. First
CWQ sera were prepared with serum-pharmacology methods. Then
the modified hypoxic chamber was designed and flushed with 5% CO2 and 95% N2 at 37 °C to induce a hypoxic environment. The effect of CWQ serum on the viability of LoVo cells was tested with MTT cytotoxicity assay. The wound model and chamber model were established to estimate the effects of CWQ serum on migration and invasion of LoVo cells. The model for cell adhesion was established to evaluate the effect of CWQ serum on LoVo cells’ adhesion. The gelatin zymography model was performed to determine the effects of CWQ serum on the activities of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). The effects of CWQ serum on the hypoxia-inducible factor 1 α (HIF-1α) nuclear translocation and the mRNA level of vascular endothelial growth factor (VEGF) in LoVo cells were determined by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses
respectively. CWQ inhibited LoVo cells’ migration based on wound healing assay. The inhibitive effect could reach about 68.00% under hypoxic culture and about 29.87% under normoxic culture when cells were treated with 10% CWQ serum for 24 h. The results from both cell invasion and adhesion assays showed that CWQ serum could dose-dependently repress the invasion of LoVo cells and inhibit cells from adhering to extra cellular matrix (ECM). Under the hypoxic culture condition
RT-PCR analysis showed that 10% CWQ serum had down-regulated the expression of VEGF by 45.87%
and the result of Western blot analysis provided further evidence. The HIF-1α amount in the nucleus of the LoVo cells was also diminished in a dose-dependent manner
as shown by the Western blot. Gel zymogram assay revealed that CWQ serum could suppress the activities of MMP-2 and MMP-9. CWQ could effectively inhibit tumor metastasis in vitro The antimetastatic effects of CWQ were associated with the inhibition of cell motility
which was evidenced by inhibition of cell invasion and adhesion. The molecular mechanisms of the inhibition of tumor invasion by CWQ were due to the reduced expression of both HIF-1α and VEGF and the suppression of MMP-2 and MMP-9 expression.
Abstract
To investigate the in vitro effects and the primary mechanisms of Changweiqing (肠胃清
CWQ) on antimetastasis and antiinvasion of hypoxic colon carcinoma cells. In addition
to provide experimental evidence for the Chinese medicinal theory of “strengthening the body’s resistance to eliminate pathogenic factors” in the treatment of colorectal cancer
including its invasion and metastasis. First
CWQ sera were prepared with serum-pharmacology methods. Then
the modified hypoxic chamber was designed and flushed with 5% CO2 and 95% N2 at 37 °C to induce a hypoxic environment. The effect of CWQ serum on the viability of LoVo cells was tested with MTT cytotoxicity assay. The wound model and chamber model were established to estimate the effects of CWQ serum on migration and invasion of LoVo cells. The model for cell adhesion was established to evaluate the effect of CWQ serum on LoVo cells’ adhesion. The gelatin zymography model was performed to determine the effects of CWQ serum on the activities of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). The effects of CWQ serum on the hypoxia-inducible factor 1 α (HIF-1α) nuclear translocation and the mRNA level of vascular endothelial growth factor (VEGF) in LoVo cells were determined by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses
respectively. CWQ inhibited LoVo cells’ migration based on wound healing assay. The inhibitive effect could reach about 68.00% under hypoxic culture and about 29.87% under normoxic culture when cells were treated with 10% CWQ serum for 24 h. The results from both cell invasion and adhesion assays showed that CWQ serum could dose-dependently repress the invasion of LoVo cells and inhibit cells from adhering to extra cellular matrix (ECM). Under the hypoxic culture condition
RT-PCR analysis showed that 10% CWQ serum had down-regulated the expression of VEGF by 45.87%
and the result of Western blot analysis provided further evidence. The HIF-1α amount in the nucleus of the LoVo cells was also diminished in a dose-dependent manner
as shown by the Western blot. Gel zymogram assay revealed that CWQ serum could suppress the activities of MMP-2 and MMP-9. CWQ could effectively inhibit tumor metastasis in vitro The antimetastatic effects of CWQ were associated with the inhibition of cell motility
which was evidenced by inhibition of cell invasion and adhesion. The molecular mechanisms of the inhibition of tumor invasion by CWQ were due to the reduced expression of both HIF-1α and VEGF and the suppression of MMP-2 and MMP-9 expression.
关键词
Changweiqingmetastasishypoxia-inducible factor 1 αmatrix metalloproteinasevascular endothelial growth factor
Keywords
Changweiqingmetastasishypoxia-inducible factor 1 αmatrix metalloproteinasevascular endothelial growth factor
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