| Objective:To explore the expression of insulin-like growth factor-1(IGF-1),insulin-like growth factor binding protein-3(IGFBP-3),matrix metalloproteinases-14(MMP-14)and urokinase-type plasminogen activator(u-PA),and to investigate their relationship in colorectal cancer tissue with clinical and pathologic factors,as so as offering a new idea to make certain the mechanism of colorectal cancer.Methods:Immunohistochemistry was used to detect the expression of IGF-1,IGFBP-3,MMP-14 and u-PA protein in normal colorectal mucosa,colorectal adenoma and colorectal cancer.Also western blot and qReal-Time PCR technology were used to detect the expression of IGF-1,IGFBP-3,MMP-14 and u-PA protein and mRNA in normal colorectal mucosa and colorectal cancer tissue.Finishing the experiments,we gathered and analyzed the data,the difference was statistically significant when P<0.05.Results:1.Results of Immunohistochemistry1.1 The expression of IGF-1,IGFBP-3,MMP-14 and u-PA protein in normal,colorectal adenoma and colorectal cancer tissuesThe expression of IGF-1,MMP-14 and u-PA protein in colorectal adenoma and colorectal cancer tissues were higher than normal colorectal mucosa(P<0.05,P<0.05,P<0.05),but the relative expression levels of IGFBP-3 protein in colorectal adenoma and colorectal cancer tissues were lower than normal colorectal mucosa(P<0.05).1.2 The relation between clinical and pathologic characteristics and the protein expression level of IGF-1,IGFBP-3,MMP-14 and u-PA in colorectal cancerIn colorectal cancer the protein expression of IGF-1,IGFBP-3,MMP-14 and u-PA was not significantly related with age,sex and differentiation of colorectal cancer(P>0.05,P>0.05,P>0.05,P>0.05).In colorectal cancer,the protein expression levels of IGF-1,MMP-14 and u-PA with muscularis propria invasion were higher than those with no muscularis propria invasion(P<0.05,P<0.05,P<0.05),but the protein expression relative levels of IGFBP-3 was different,it was not correlated with colorectal cancer muscularis propria invasion(P>0.05).The protein expression relative levels of IGF-1,MMP-14 and u-PA in colorectal cancer with lymph node metastasis were higher than with no lymph node metastasis(P<0.05,P<0.05,P<0.05).Conversely,the protein expression levels of IGFBP-3 was lower with lymph node metastasis than with no lymph node metastasis(P<0.05).In colorectal cancer,III and IV stage of TNM the protein expression levels of IGF-1,MMP-14 and u-PA were higher than those I and II stage of TNM(P<0.05,P<0.05,P<0.05),but the protein expression levels of IGFBP-3 was higher in I and II stage of TNM than III and IV stage of TNM in colorectal cancer(P>0.05).2.The results of Western Blot2.1 Protein expression of IGF-1,IGFBP-3,MMP-14 and u-PA in normal tissues and colorectal cancerThe relative expression levels of IGF-1,MMP-14 and u-PA protein in colorectal cancer were higher than in normal colorectal mucosa(P<0.05,P<0.05,P<0.05),yet the protein expression levels of IGFBP-3 was higher in normal colorectal mucosa than colorectal cancer(P<0.05).2.2 The relation between clinical and pathological characteristics and the protein expression of IGF-1,IGFBP-3,MMP-14 and u-PA in colorectal cancerThe protein expression of IGF-1,IGFBP-3,MMP-14 and u-PA in colorectal cancer was not significantly correlated with age,sex and differentiation of colorectal cancer of patients(P>0.05,P>0.05,P>0.05,P>0.05).In colorectal cancer tissues the expression relative levels of IGF-1,MMP-14 and u-PA protein were higher with lymph node metastasis than with no lymph node metastasis(P<0.05,P<0.05,P<0.05),but the protein expression levels of IGFBP-3 was higher with no lymph node metastasis than with lymph node metastasis(P<0.05).The relative protein expression levels of IGF-1,MMP-14 and u-PA were lower in I and II stage of TNM in colorectal cancer than those III and IV stage of TNM(P<0.05,P<0.05,P<0.05),yet in I and II stage of TNM in colorectal cancer the protein expression levels of IGFBP-3 was higher than III and IV stage of TNM(P<0.05).In colorectal cancer the protein expression relative levels of IGF-1,MMP-14 and u-PA were higher with muscularis propria invasion than with no muscularis propria invasion(P<0.05,P<0.05,P<0.05),but the protein expression levels of IGFBP-3 was not related with colorectal cancer muscularis propria invasion(P>0.05).3.Results of qReal-Time PCR3.1 The mRNA expression of IGF-1,IGFBP-3,MMP-14 and u-PA in normal and colorectal carcinoma tissuesIn colorectal cancer,the mRNA expression levels of IGFBP-3 was lower than in normal colorectal tissues(P<0.05),but the expression levels of IGF-1,MMP-14 and u-PA mRNA was higher in colorectal cancer than in normal colorectal tissues(P<0.05,P<0.05,P<0.05).3.2 The relationship between mRNA expression of IGF-1,IGFBP-3,MMP-14,u-PA and clinicopathological features in colorectal carcinomaIn colorectal cancer the mRNA expression of IGF-1,IGFBP-3,MMP-14 and u-PA was not related with age,sex and differentiation of colorectal cancer(P>0.05,P>0.05,P>0.05,P>0.05).The relative expression levels of IGF-1,MMP-14 and u-PA mRNA were higher in III and IV stage of TNM in colorectal cancer than those I and II stage of TNM(P<0.05,P<0.05,P<0.05),yet in I and II stage of TNM in colorectal cancer the mRNA expression levels of IGFBP-3 was higher than III and IV stage of TNM(P<0.05).With no muscularis propria invasion in colorectal carcinoma tissues the relative mRNA expression levels of IGF-1,MMP-14 and u-PA were lower than those with muscularis propria invasion(P<0.05,P<0.05,P<0.05),yet the mRNA expression levels of IGFBP-3 was not correlated with muscularis propria invasion in colorectal cancer(P>0.05).In colorectal carcinoma tissues the relative mRNA expression levels of IGF-1,MMP-14 and u-PA were higher with lymph node metastasis than those with no lymph node metastasis(P<0.05,P<0.05,P<0.05),but with no lymph node metastasis the relative mRNA expression levels of IGFBP-3 was higher than with lymph node metastasis in colorectal cancer(P<0.05).4.Correlation of IGF-1,IGFBP-3,MMP-14 and u-PA in colorectal cancerImmunohistochemistry,Western Blot and real-time fluorescence quantitative PCR experiments showed that the expression of IGFBP-3 was negatively correlated with the expression of IGF-1,MMP-14 and u-PA,respectively(P<0.05,P<0.05,P<0.05);In the Western Blot experiment,the expression of IGF-1,MMP-14 and u-PA were expressed.There was a positive correlation between them(P<0.05,P<0.05,P<0.05).In the real-time fluorescence quantitative PCR assay,MMP-14 was positively correlated with the expression of IGF-1 and u-PA,respectively(P<0.05,P< 0.05).Conclusion:1.From the normal intestinal mucosa group to the adenoma group and then to the cancer,the expression of IGFBP-3 was down-regulated,and the expression levels of IGF-1,MMP-14,and u-PA increased in turn,suggesting that the four involved in the occurrence of colorectal cancer.Overexpression of IGF-1,MMP-14,u-PA,and low expression of IGFBP-3 promote tumorigenesis.2.The expression of IGF-1,MMP-14,and u-PA in colorectal cancer tissues with infiltrative depth invading into muscularis propria,lymph node metastasis,and TNM stage III+IV was higher than that of infiltration and intrinsic muscle Stratum,no lymph node metastasis and TNM staging are cancerous tissues at stage I+II;while the expression of IGFBP-3 in lymph node metastasis,and TNM stage III+IV colorectal cancer tissues is lower than lymph node metastasis and TNM I + II stage cancer tissue,suggesting that IGF-1,MMP-14,u-PA and IGFBP-3 participate in the invasion and metastasis of colorectal cancer tissue.3.IGFBP-3 may play an important role in the development of colorectal cancer through regulating the expression of IGF-1,MMP-14 and u-PA.Increasing the expression of IGFBP-3 may improve the patient's treatment and prognosis.4.The detection of IGF-1,IGFBP-3,MMP-14 and u-PA alone or in combination is expected to be one of the molecular targeted therapies.It can be used not only in colorectal cancer but also in the diagnosis,treatment,and prognosis of other systemic tumors. |
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