【Abstract】ObjectiveTo study the expressions of matrix metalloproteinase 2 (MMP2) and carbohydrate antigen 50 (CA50) in colorectal carcinoma, cancer-adjacent mucosa （2 cm from the nether edge of tumor）, cancerdistant mucosa （5 cm from the nether edge of tumor） and normal colorectal mucosa, and to elucidate their effects on the development of colorectal carcinoma. MethodsThe expressions of MMP2 and CA50 were detected immunohistochemically in 40 cases of colorectal carcinoma, cancer-adjacent mucosa, cancer-distant mucosa and 10 cases of normal colorectal mucosa. Results①The expression intensity and positive rates of MMP-2 and CA50 increased significantly in turn by normal mucosa, cancer-distant mucosa, cancer-adjacent mucosa and colorectal carcinoma. ②The expression of MMP2 was correlated with CA50 in colorectal carcinoma. ③The expression of CA50 in colorectal carcinoma was closely associated with tumor differentiation, and the expression of MMP2 in colorectal carcinoma was closely associated with differentiation and Dukes stages as well. ConclusionOver expression of MMP2 facilitates the malignant progress of colorectal carcinoma; CA50 is a reliable marker of malignance in colorectal carcinoma; CA50 and MMP2 may have synergetic effects on the development of colorectal carcinoma.
Objective To investigate the effects of X-ray dose on the expressions of microRNA-221 （miR-221） and phosphatase and a tensin homolog deleted from chromosome10 （PTEN） in human colorectal carcinoma （CRC） cells. Methods Human CRC-derived cell line， Caco2， was cultured conventionally. The cells were divided into five groups and exposed to different doses of X-ray （0， 2， 4， 6， and 8 Gy） respectively. The total RNA and protein of the Caco2 cells were extracted after irradiation， and the miR-221 and PTEN mRNA expressions were detected by real-time RT-PCR.Moreover， the protein alteration of PTEN in Caco2 cells was detected by Western-blot analysis. Results The radiation dose of X-ray significantly affected the expressions of miR-221 and PTEN protein in human Caco2 cells in a dose-depen-dent manner. Moreover， the miR-221 expression level was up-regulated gradually with the increase of irradiation dose， on the contrary， the PTEN protein expression level was down-regulated gradually （P＜0.01）. Conclusion The radiation dose can affect the miR-221 and PTEN protein expression pattern in CRC cells.
To study the significance of T-lymphocytes rDNA transcription activity in diagnosis, differential diagnosis, therapeutical effect and evaluation of treatment for colorectal carcinoma, 59 cases of colorectal carcinoma, 20 cases of colorectal inflammatory disease and 9 volunteers were choosen to detect the T-lymphocyte rDNA transcription activity of peripheral blood T-lymphocyte by cell culture and CMIAS008 image analysis system of Ag-NOR. Results: T-lymphocytes rDNA transcription activity was decreased obviously in colorectal inflammatory patients. Compared with control group, both group showed markedly statistical difference (Ｐ＜0.01). Tlymphocytes rDNA transcription activity increased gradually to normal groups after operation and chemical treatment for colorectal carcinoma patients; but it decreased for recurrent patients three years after operation. Conclusions: The detection of T-lymphocytes transcription activity can be used as a differential criterion for colorectal carcinoma and colorectal inflammatory disease, meanwhile it also can be used as a reference criterion for evaluation of treatment and supervision of tumor recurrence.
Objective To investigate the prevalence of cellular FLICE-like inhibitory protein (cFLIP) alterations in colorectal cancer and their possible implications for the progression of colorectal cancer. Methods The long type cFLIP (cFLIPL) was examined in 43 colorectal cancer specimens and the matched normal colorectal specimens by immunohistochemistry. Immunohistochemical staining for cFLIPL was assessed on an arbitrary semi-quantitative scoring system. Stained cells were counted under the magnification field (×100) and at least 20 fields per case were examined. Fields with non-stained cells were scored 0; fields with stained cells less than 5% were scored 1; fields with stained cells from 5% to 25% were scored 2; fields with stained cells between 26% and 50% were scored 3; and fields with stained cells more than 50% were scored 4. According to the above schedule， a mean score of each specimen was calculated. Results cFLIPL protein expressions of variable intensity and extent were detected in the normal colorectal mucosa and adenocarcinomas. cFLIPL was mainly expressed in the cytoplasma. The positive cells were distributed in diffuse manner. The mean expression score in normal mucosa ranged from 0 to 2.95 (mean score: 1.55±0.86); 4.7%(2/43) of all cases were unstained and 20.9%(9/43) showed a weakly stained pattern (0＜mean score≤1); 74.4%(32/43) of all cases were positively stained (1.00＜mean score≤2.95), respectively. cFLIPLprotein was expressed in all adenocarcinomas and the score ranged from 0.80-4.00 (mean score: 3.06±0.75); 62.8% (27/43) of the tumors examined were predominantly cFLIPL positive (mean score ＞3), 34.9%(15/43) of the tumors were cFLIPLpositive (1＜mean score ≤3) and only one case was cFLIPL weakly positive (score: 0.80). 72.1% (31/43) of adenocarcinomas expressed cFLIPLmore intensely and extensively than matched normal colonic mucosa. cFLIPL expression of colorectal cancer was significantly higher than that of matched normal mucosa, which was statistically significant (P＜0.01). The extent of cFLIPL expression in tumors was independent of Dukes stage, tumor stage, gross type of tumor, histological type, or lymph and hepatic metastasis. Conclusion The expression level of cFLIPL in adenocarcinomas is much higher than that in matched normal mucosa. Overexpression of cFLIPL is a tumor-specific phenomenon, which can provide a selective growth advantage for colorectal cancer cells to escape from death receptor-mediated apoptosis.
ObjectiveTo evaluate the effect of neoadjuvant chemotherapy and find the mechanism of multidrug resistance. MethodsTwenty patients with gastric cancer and 31 patients with colorectal cancer underwent neoadjuvant chemotherapy and then operations. The preoperative specimens were stained by immunohistochemical techniques for testing p53,multidrug resistanceassociated protein (MRP), glutathione S transferase（GST）, telomerase. Resection specimens were evaluated for chemotherapy effect by routine histology; at the same time, the postoperative morbidity and mortality were observed. ResultsIn 51 patients, the response rate of neoadjuvant chemotherapy was 27.45%(14/51),so multidrug resistance was a kind of common phenomena in gastrointestinal carcinomas. The postoperative morbidity was 15.69%(8/15), the main operation complication was infection,the mortality was 1.96%(1/51),only one person died from severe infection.The expression rate of p53, MRP, GST, telomerase was 58.0%,51.0%,66.7%,74.0%respectively, the location of p53 was at cell nucleus,location of MRP,GST was at cell memberane and cytoplasm,location of telomerase was at cytoplasm.The response rate had nothing to do with age, sex and metastasis. But it was related with p53 and telomerase expression. ConclusionNeoadjuvant chemotherapy is an effective, safe therapy. But the rate of drug resistance is high in gastrointestinal carcinomas, and the response rate is related to p53, telomerase expression.
Objective To evaluate the relationship between leptin level in serum and clinicopathologic features of colorectal cancer. Methods ABC-ELLSA was used to detect the leptin level in 30 cases of colorectal cancer without dystrophy (cancer group) and 24 normal controls (control group). The expressions of K-ras， p53， adenomatous polyposis coli （APC） gene and delete in colorectal carcinoma gene (DCC) mRNA of the tumor were examined by RT-PCR, the levels of serum CEA and CA19-9, and other clinicopathologic features were also recorded. Results The leptin level in cancer group 〔（3.53±1.72） μg/L〕 was higher than that in control group 〔（2.27±1.01） μg/L〕, P＜0.05, and the difference was independent on gender. There were no significant differences of leptin level in different tumor stages and different tumor location （Pgt;0.05）. Leptin level of poorly differentiated tumor was obviously lower than that of well differentiated and moderately differentiated tumor （P＜0.05）. There were no associations between leptin level and the levels of CEA and CA19-9, likewise there were no associations between leptin level and the expressions of K-ras, p53, APC and DCC in tumor （Pgt;0.05）. Conclusion The leptin level of colorectal cancer patient is higher than that of normal person, which is affected by the differentiation of tumor. But there are no significant correlations between the level of leptin in serum and TNM stage, tumor location, tumor markers of serum, K-ras, p53, APC or DCC in tumor.
ObjectiveTo detect expressions of epidermal growth factor receptor (EGFR), BRAF, and K-Ras genes in colorectal carcinoma tissues and explore pathogenesis in colorectal carcinoma. MethodThe expressions of EGFR, BRAF, and K-Ras genes were detected in these 136 colorectal carcinoma tissues and their corresponding adjacent normal colorectal tissues by immunohistochemistry. ResultsThe expressions of EGFR and BRAF in the colorectal carcinoma tissues were significantly higher than those in their corresponding adjacent normal colorectal tissues (P<0.05), but the expression of K-Ras had no significant difference between these two tissues (P>0.05). The expression of EGFR gene was related to the TNM stage, lymphatic metastasis, or degree of differentiation. The expression of BRAF gene was related to the TNM stage. The expression of K-Ras gene wasn,t related to the TNM stage, lymphatic metastasis, or degree of differentiation. The correlation analysis results showed that there was no relation among the EGFR, K-Ras, or BRAF expression. ConclusionsUp-regulated of EGFR and BRAF gene expressions might be related to development of colorectal carcinoma, and role of K-Ras is unclear. Anti EGFR and BRAF target therapy might be benefited for patients with colorectal carcinoma.
Objective To investigate the level of cell adhesion molecule sialyl-LeX expression in colorectal carcinoma and its relation with carcinogenesis, differentiation, metastasis and prognosis. Methods Sialyl -LeX expression and its optical density in colorectal carcinoma (n=90) and remote normal mucosa (n=30) were quantitatively studied with microwave-LSAB immunohistochemical method combined with image analysis technique. Fifty-three patients were followed up. Results The weaker staining in remote normal colorectal mucosa was observed in very limited parts of some deep crypts. Positive rate of sialyl-LeX expression was only 16.7%(5/30). The positive expression of sialyl-LeX was observed in 83 of 90 patients with colorectal carcinoma(92.2%). The apical cytoplasma of cancer tubules, the luminal contents, and the cytoplasma of the cancer cells were bly stained. The mean integral optical density of sialyl-LeX positive cell in poorly differentiated adenocarcinoma was significantly higher than that in highly differentiated and mucinous ones (Plt;0.01). It was markedly higher in patients with positive lymphatic nodes than that in negative ones (Plt;0.01). With followed-up for longer than 5 years, it was much lower in the alive cases than that in the dead (Plt;0.01). Conclusion These findings indicate that changes of sialyl-LeX expression and its optical density is related to carcinogenesis, differentiation, invasion and metastasis of colorectal carcinoma. It may be a good predicter for the prognosis of patients with colorectal carcinoma.
Objective To detect expression of CK20 mRNA in peripheral blood of patients with colorectal carcinoma and its clinical significance. Methods Using the reverse transcriptase-polymerase chain reaction (RT-PCR),CK20 mRNA expression was examined in peripheral blood from 42 patients with colorectal carcinoma before and after operation, 20 healthy volunteers, 20 fresh colorectal carcinoma samples. Results The positive expression rates of CK20 mRNA were 45.24%(19/42) and 33.33%(14/42) before and after operation in 42 colorectal carcinoma patients respectively. All 20 fresh colorectal carcinoma samples revealed expression of CK20 mRNA, but the 20 normal blood samples did not. Conclusion The detection of CK20 mRNA in peripheral blood is helpful to early diagnose, assess the prognosis and make a correct treatment of colorectal carcinoma.
ObjectiveTo evaluate the significance and effect for surgery of Dukes D stage colorectal carcinoma. MethodsEightytwo cases of Dukes D stage colorectal carcinoma who underwent surgery from 1991 to 1998 were analyzed retrospectively. ResultsTwentyone patients experienced extended resections,29 patients palliative resection, 19 patients sideside anastomosis and 13 patients laparotomy and biopsy of the tumor. Patients with extended resections had significantly long survival time, quality of life in these patients were excellent in 71.4%, good in 23.8%, and fair in 4.8%; whereas quality of life in the group with palliative resections were excellent in 62.1%, good in 17.2%, fair in 13.8%, and poor in 6.9%. The other two groups had a bad postoperative effect, short survival time and poor quality of life. ConclusionIf the patients with Dukes D stage colorectal carcinoma have operative indications,surgical therapy should be actively performed and the postoperative survival rate and quality of life can be improved.