| ObjectiveTo investigate the expression of CD44and Ki-67(the cell cycle marker) and their clinical pathology significance in colorectal carcinomas. To explore the location, quantity and cell cycle status of CD44positive cancer stem cells. To observe the distribution, quantity, morphological characteristics of the CD44+/Ki-67-cancer stem cells in colorectal carcinoma tissue and to investigate its value as cancer stem cells marker. Furthermore, to observe the morphological characteristics and quantity of the CD44+/Ki-67-cells in SW620cell line of human colorectal carcinomas, to compare the expression with solid tumor tissue of colorectal carcinoma and to confirm the value of CD44-/Ki-67-phenotype as a marker of cancer stem cells of colorectal carcinoma in cytology level.MethodsThe expression of CD44and Ki-67were detected in10cases of normal mucosa tissues,15cases of adenoma,59cases of colorectal carcinoma and SW620cell lines of human colorectal carcinoma and observed location, quantity and clinical pathology relationship of the CD44+/Ki-67-cancer stem cell by tissue chip (tissue microarray) technology, immunohistochemical staining and double immunohistochemical staining technology.Results1. The expression of CD44, Ki-67and CD44+/Ki-67" in normal tissue, adenoma and colorectal carcinomaIn normal mucosa, the expression of CD44protein was negative, and low rate in adenoma; In carcinoma tissue, the expression of CD44in the intensity and number was different accounting for0.01%-32%; The positive cells had two kinds of cells form, which one was a larger nucleus, round or oval, obvious nucleoli; Another was a small nuclei, round or oval. In normal mucosa, Ki-67was less than3%, mostly locating in the base membrane of bowel mucosa. In adenoma, Ki-67was more than5%and highly expression in nuclear division abound area. In the carcinoma tissue, the quantity of Ki-67accounted for1%-70%and the average was24.98%. But the Ki-67" took account for30%-97%and the average of75.02%. The average quantity of the CD44+Ki-67-cancer stem cells was5.82%, which was small nuclei, round or oval, similar to a lymphocytes, and mainly distributed along the side of the gland basement wall of the glands or side wall.2. The expression and clinical pathology significance of CD44, Ki-67and CD44-/Ki-67-in colorectal carcinomaThere was significant difference in the expression of CD44in normal mucosa and adenoma comparing with the carcinoma tissue (P<0.05); There was significant difference between the expression of Ki-67in normal mucosa and the carcinoma tissue (P<0.01) and also significant difference between the adenoma and the carcinoma tissue (P<0.05); But there was no difference in the normal mucosa and adenomas (P>0.05). In colorectal carcinoma, the expression of CD44-/Ki-67-was positively relevant with infiltration depth, lymph node metastasis and Duck s stage of carcinoma (P<0.05).3. The expression and relationship of CD44and Ki-67in colorectal carcinomaIn Ki-67" carcinoma cell, there was significant difference between CD44+cells accounted for67%and CD44-cells occupied for33%(P<0.05). Most colorectal carcinoma CD44-cancer stem cells were in static or GO and a few were in the cell cycle of the Gl, S, G2and M period.4. The expression of CD44and Ki-67in colorectal carcinoma cell lines SW620In colorectal carcinoma cell lines SW620, CD44+were an average of7.56%, Ki-67+were91.94%, Ki-67-were8.06%, CD44+Ki-67-cancer stem cells were5.68%. CD44+Ki-67-cancer stem cells mainly expressed in small round cells and mono-polar cells and less in other cells.Conclusion1. In colorectal carcinoma, CD44+Ki-67-cancer stem cell mainly distribute along the basement side or common wall side of the glands, which with a small nuclei, round or oval and resembling to lymphocyte.2. In colorectal carcinoma, most of CD44+cancer stem cells situate in the stage of GO and a few in the stage of G1, S, G2and M of cell cycle.3. In human colorectal carcinoma cell lines SW620, CD44+/Ki-67" cancer stem cells are similar to the solid tumor tissues in the morphological characteristics, quantity and so on.4. The CD44+/Ki-67-phenotype is a valuable marker of cancer stem cells and effective index for recurrence, metastasis and prognosis of colorectal carcinoma. |
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