RESUMO
BACKGROUND:DCLK1 is a transmembrane microtubule-associated kinase in neurons after mitotic division, which may be the intestinal cancer stem cel marker. OBJECTIVE:To observe the expression and pathological significance of DCLK1 and Ki67 in colorectal cancer. METHODS: Expression of Ki67 and DCLK1 in 150 cases of colorectal cancer tissues was detected by immunohistochemical method in contrast to normal colorectal mucosa, para-carcinoma tissue, and adenoma tissue. RESULTS AND CONCLUSION:The expression rates of DCLK1 and Ki67 were 36.7% and 34.7% in cancer tissues, respectively, both of which were significantly higher than those in normal colorectal mucosa and adenoma. The expression of DCLK1 was associated with the location, depth of invasion, lymph node metastasis (P < 0.05), while the expression of Ki67 was just associated with the depth of invasion (P < 0.05). There was a negative correlation between the expression of DCLK1 and Ki67 (r=-0.460,P=0.000). The count of DCLK1+/Ki67-cels was about 2.01% in colorectal cancer tissues, and these cels mainly distributed at the bottom of intestinal mucosa base and common duct wal. DCLK1+/Ki67- cels were oval, the nuclei were large and deep-stained with prominent nucleolus, and there was rare nuclear fission and less cytoplasm. From the aspects of cel number, location, and cel morphology, DCLK1+/Ki67- cels are in line with the characteristics of cancer stem cels; therefore, DCLK1+/Ki67-can be used as a cancer stem cel marker of colorectal cancer.
RESUMO
Objective :To construct hepatocarcinoma specific IL-1? anti-sense RNA expression vector and to explore its effect on the growth of implanted hepatocarcinoma H22 cells in mice and the possible mechanism. Methods:Murine IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 under the regulation of minimal alpha-feto protein (AFP) promoter and CMV enhancer were constructed,and further verified by PCR,restriction endonuclease analysis and DNA sequencing. H22 cells transfected with pafpIRES2-antiIL-1? 1 or pafpIRES2-antiIL-1? 2 were divided into 3 groups:H22/mock,H22/antiIL-1?1 and H22/antiIL-1?2 group. Expression of IL-1? was detected by RT-PCR. Transfected H22 cells were subcutaneously injected into mice to establish tumor implanted mouse model. Tumor volume was measured; the cytotocixity of spleen NK against H22 cells was detected by MTT. Results:Hepatocarcinoma specific IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 were successfully constructed and were verified by PCR,restriction endonuclease analysis and DNA sequencing. IL-1? expression in H22 cells was down-regulated after transfected with IL-1? anti-sense RNA expression vectors,especially with the pafpIRES2-antiIL-1?2 vector. Hepatocarcinoma cells implanted mouse model was successfully established. Tumor volume and growth of tumor in H22/antiIL-1?2 mice was obviously smaller than that in H22/mock mice,and the cytotocixity of spleen NK against H22 cells in H22/antiIL-1?1 and H22/antiIL-1?2 mice was also greatly enhanced. Conclusion:Hepatocarcinoma specific IL-1? anti-sense RNA expression vector pafpIRES2-antiIL-1? was successfully constructed. It effectively inhibits the growth of implanted hepatocarcinoma in mice probably through specifically blocking expression of IL-1? and increasing cytotocixity of spleen NK.
