ObjectiveTo investigate healing of rat colonic anastomoses after early postoperative intraperitoneal chemotherapy (EPIC).MethodsFortyfive Wistar rats with colonic anastomoses were divided randomly into 3 groups (15 rats each). From postoperative 1 day to 5 day, rats were injected with normal saline (NS) to the peritoneal cavity with 20 ml/(kg·d) for the NS group; 5Fu with 20 mg/(kg·d) for the 5Fu group; 5Fu with 20 mg/(kg·d) and leucovorin with 10 mg/(kg·d) for the 5Fu+LV group. On the 7th postoperative day, rats were killed and the anastomoses were evaluated whether anastomotic complications (leakage or dehiscence) occurred, the anastomotic bursting pressure (ABP) and hydroxypoline content (HPC) were measured. ResultsIn the NS group, 1 rat had incision dehiscence, another one had anastomostic leakage with but no death. In the 5Fu+LV group, 2 rats showed anastomotic leakage and 1 death. On the 7th postoperative day, the ABP in NS, 5Fu and 5Fu+LV groups were (169.1±32.6) mm Hg, (116.8±25.5) mm Hg and (154.9±31.2) mm Hg respectively; the HPC was (1.54±0.28) μg/mg, (0.9±0.33) μg/mg and (1.24±0.29) μg/mg respectively. Both the ABP and HPC, in the NS group were much significantly higher than in 5Fu group (P＜0.01). Both the ABP and HPC in the 5Fu+LV group were significantly higher than which in the 5Fu group (P＜0.05).ConclusionEPIC with 5Fu significantly impairs healing of the colonic anastomosis. 5Fu combined with LV for EPIC might reduce this inhibition to the process of the anastomotic healing.
Objective To study the relationship between blood metastasis of colorectal cancer and cancer metastasis related factors.MethodsCK20 mRNA in peripheral blood was investigated by reverse transcription polymerase chain reaction (RTPCR) and proteins of CD44v6 and p53 in cancer tissues were examined by immunohistochemical in 50 cases of colorectal neoplasm. ResultsThe results showed that the positive rates of peripheral blood micrometastasis of colorectal cancer were 68%. It escalated along with the rising of the Dukes stage, the rates in Dukes C and D stage were significantly higher than that in Dukes A and B stage. The positive rates of CD44v6,p53 expression in colorectal cancer were 74% and 62% respectively. The positive rates of CD44v6 and p53 in Dukes A and B stage were significantly lower than those in Dukes C and D stage,in peripheral blood and colorectal cancer micrometastasispositive group were significantly higher than that in the micrometastasisnegative group. CK20 mRNA was significantly correlated with expressions of CD44v6 and p53 in cancer tissues. Conclusion The detection of CK20 mRNA in blood before operation and after operation examination of CD44v6 and p53 in cancer tissues are helpful for prediction of blood metastasis of colorectal neoplasm and postoperative treatment.
ObjectiveTo investigate the expression and clinical significance of cyclin dependent kinase 8 (CDK8) and transforming growth factor-β1 (TGF-β1), and their correlation in human colorectal cancer tissues. MethodsThe CDK8 and TGF-β1 mRNA expressions were examined by using real-time quantitative polymerase chain reaction and the protein expresssions of them were detected by immunohistochemistry on a cohort of human colorectal cancer tissues (n=40) and corresponding adjacent tissues (n=40). The correlation between CDK8 and TGF-β1 was also analyzed. ResultsThe expression levels of CDK8 mRNA and TGF-β1 mRNA in colorectal cancer were dramatically increased compared with the corresponding adjacent tissues (P < 0.05). The expression level of CDK8 mRNA was significantly associated with lymphnode metastasis, depth of invasion, and colorectal cancer stage (P < 0.05). However, the expression level of CDK8 mRNA was no correlation with the age, gender of patients, tumor size and differentiation of colorectal cancer in this study. The expression level of TGF-β1 mRNA was significantly associated with lymphnode metastasis, depth of invasion, tumor size and colorectal cancer stage (P < 0.05). However, the expression level of TGF-β1 mRNA was no correlation with the age, gender of patients and differentiation of colorectal cancer. The expressions of CDK8 mRNA and TGF-β1 mRNA in colorectal cancer was positively correlated (r=0.387, P=0.048). ConclusionsThe upregulation of CDK8 and TGF-β1 expression may be related to the occurrence and development of colorectal cancer, and the two may have a synergistic effect, which may be related to the biological behavior and prognosis of colorectal cancer.
The model of transplanted colonic SW480 cell line carcinoma in gymnomouse body was set up to observe the effect of octapeptide somatostatin (SMS 201-995,SMS) on the transplanted carcinoma and elucidate its mechanism. Results: the volume, weight, DNA and protein content in carcinoma cell, cell amount and proliferation index of S and G2M phase in SMS group and SMS+PG (pentagastrin) group were markedly lower than those in PG group and control group, those of PG group were markedly higher than those in control group.The cell amount of G0/G1 phase in SMS group and SMS+PG group was markedly higher than that in PG group and control group, and that of PG group was markedly lower than that in control group.All these suggested that somatostatin could not only inhibit the growth of transplanted human colonic SW480 cell line carcinoma directly but also inhibit the growthpromoting effect of gastrin on the transplanted carcinoma.The mechanism might be that somatostatin inhibit the synthesis of cAMP, DNA and protein in carcinoma cells, then inhibit the cell growing from G0/G1 phase to S and G2M phases.Our study might provide experimental basis for the homonotherapy with analogue of somatostatin in patients with large intestine carcinoma.
Objective To study the effects of long term application of cathartics on electromyography of rat colon, and to explore the role of interstitial cells of Cajal (ICC) in it. Methods Colonic slow waves of the rat was examined after 3-month feeding of phenolphthalein, and ICC in myenteric plexus was observed by ZIO method, and ultrastructure changes of nerves and ICC was observed. Results The frequency of slow waves of cathartic colon was reduced significantly(Ｐ＜0.05). The distribution of ICC in myenteric plexus was uneven, and the processes were mussily connected each other. Vacuolar degeneration of axon and ICC-like cells was revealed by electron microscope in myenteric plexus of cathartic colon. Conclusion Long term application of phenolphthalein could reduce the frequency of colonic slow waves, and the possible mechanism was degeneration of ICC and myenteric plexus nerves.
Objective To investigate the mechanism and clinical significance of vincristine (VCR) inhibiting gastrinproliferation effects on human colon cell line SW480. Methods Effects of VCR on the viable cell count (A value), myoinositol triphosphate (IP3, CPM value), 〔Ca2+〕i and protein kinase C (PKC) activity of human colon cell line SW480 were evaluated in vitro by MTT assay,3Hmyoinositol incorporation, fluorescence measurements and γ-32P-ATP incorporation.Results A value of VCR+PG group was lower than that of PG or control group (Ｐ＜0.01 vs control, Ｐ＜0.01 vs PG). The concentration of IP3 or 〔Ca2+〕i in VCR+PG group was lower than that in PG group (Ｐ＜0.01 vs PG); and the PKC activity of membrane was lower than that in PG group (Ｐ＜0.05 vs PG, Ｐ＞0.05 vs control). Conclusion Effects of vincristine may be through the phosphoinositide signaling pathway on gastrinstimulating cell proliferation in human colon cell line SW480. It has provided an experimental evidence for antisignaling therapy for patients with colon cancer.
Objective To evaluate the significance of serum colon cancer-specific antigen-2 （CCSA-2） in diagnosisof colorectal cancer （CRC）. Methods By using ELISA method， the serum CCSA-2 was measured from 105 patients with 5 kinds of diseases， including CRC， gastric cancer， inguinal hernia， acute appendicitis， and breast cancer， who weretreated in our hospital from Jul. to Dec. in 2008， and 20 health donors were enrolled in addition. The blood samples were collected on 3 days before surgery， but blood samples from patients with acute appendicitis were collected before emergencysurgery， blood samples of health donors were collected on 1 day before ELISA test. Results The level of serum CCSA-2 in CRC patients was （99.27±6.25） μg/L， which was significantly higher than those of other patients and health individuals〔（53.58±2.73） μg/L， t=48.29， P=0.000〕. Serum CCSA-2 at a cutoff of 73.96μg/L had a sensitivity of 100% （95% CI：100%-100%） and a specificity of 100% （95% CI：100%-100%） in separating CRC populations from all other indivi-duals by using receiver operator characteristic curve （ROC） analysis. As compared with carcinoembryonic antigen （CEA） and CA19-9， the serum CCSA-2 assay （at a cutoff of 73.96μg/L） was significantly more sensitive than CEA and CA19-9 assay in CRC detection （P＜0.01）. Serum CCSA-2 was not related with patients’ gender （P=0.81）， age （P=0.59）， TNM stage （P=0.85）， Dukes stage （P=0.63）， nuclear grade （P=0.44）， as well as expressions of multidrug resistance associated protein （P=0.33）， P-glycoprotein （P=0.72）， and topoisomeraseⅡ（P=0.95）， but higher in patients with colon cancer than those of patients with rectal cancer （P=0.02）. Conclusion Serum CCSA-2 may be a useful biomarker in diagnosis of CRC， and it may be only related to tumorigenesis， but is irrelated to tumor progression and chemotherapy.
ObjectiveTo explore the influence mechanism of proliferation and invasion in colon cancer cell after silence of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene. MethodsRT-PCR or Western blot method was used to detect the expression of PTEN mRNA or protein among four colon cancer cell lines (HT-29, WiDr, CaCo-2, and Colo320 cell lines). small interfering RNA (siRNA) was used to synthetize PTEN siRNA and transfect it into colon cancer cells. The expression of PTEN protein after transfecting was detected by Western blot. WsT-1 and invasion assay were used to examine the effects of PTEN siRNA silence on proliferation and invasion in colon cancer cells. ResultsPTEN mRNA and protein were expressed in all the four colon cancer cell lines. After PTEN siRNA transfected into the colon cancer cells, the expressions of PTEN proteins were inhibited in all the four colon cancer cell lines (P < 0.01), and the proliferation and invasion of colon cancer cells were enhanced significantly (P < 0.01). ConclusionsPTEN siRNA play an important role in metastasis process of colon cancer via enhanced its proliferation and invasion. Therefore, the understanding biologic mechanisms for regulation of PTEN might enable better molecular target therapy of treating the colon cancer patients with metastasis.
ObjectiveTo detect 5-FU concentration and investigate the changes of pathology， and Ki-67 protein expression after intraoperative regional chemotherapy (RC) for colon cancer. MethodsAll the patients were randomized into two groups: RC group (n=20), received intraoperational RC with 100 ml physiological saline contained 5-FU (15 mg/kg) and camptothecine (0.06 mg/kg)； control group (n=20), saline alone. The samples from portal vein blood, peripheral blood, peritoneal fluid， and peri-cancerous tissues in RC group were taken to detect the 5-FU concentration by high performance liquid chromatography (HPLC), respectively at 2, 5, 10, 20, 30, and 60 minutes after treatment. The pathological changes were observed and Ki-67 protein expressions were examined by immunohistochemical staining for all the cancer tissues postoperatively in two groups. ResultsPeak concentration of 5-FU appeared at 2 min after treatment, and decreased gradually. 5-FU concentration in peritoneal fluid was the highest, and the lowest in the peripheral blood (Plt;0.01). In RC group， light karyopyknosis, nuclear swelling, and coagulative necrosis of cancer cells, and light intercellular substance hydropsia, inflammatory cells invasion were observed under light microscopic examination; light vasculitis presented also in five cases. Nuclear swelling, heterochromatin agglutination, perinuclear gap expansion, mitochondrial swelling, endoplasmic reticulum expansion, and Golgi complex expansion were observed with transmission electron microscope. Ki-67 protein expression of colon cance tissues in RC group was lower than that in control group （Plt;0.05）. Conclusions Intraoperative RC for colon cancer may sustain a high concentration of chemotherapy drugs in peritoneal fluid and portal vein blood, and alter histopathological morphology of cancer cells, and suppress Ki-67 protein expression. So, intraoperative RC may play an important role in preventing intraoperative spreading and postoperative recurrence of colon cancer.
Objective To study the feasibility and curative effect of laparoscopic vs. open radical rectectomy and colectomy for colorectal cancer. Methods Sixty-two cases who underwent laparoscopic operation (17， 2， 10， 23， 9 and 1 case underwent radical right colectomy， radical transverse colectomy， radical left colectomy， Dixon， Miles and Hartmann operation respectively) and 78 cases who underwent open operation (17， 4， 11， 27， 18 and 1 case underwent radical right colectomy， radical transverse colectomy， radical left colectomy， Dixon， Miles and Hartmann operation respectively) in our department from Aug. 2001 to Jun. 2008 were included. The clinical data of patients in two groups were compared. Results There were no severe complications and death occurred in both groups and 4 cases in laparoscopic group were converted to open operation during the procedure. The mean operation time of laparoscopic group and open group were (230.6±23.5) min and (145.5±17.6) min respectively, there was a statistical difference between them (P＜0.01). The intra-operative blood loss of laparoscopic group was obviously less than that in open group 〔(135.5±22.5) ml vs. (300.6±34.5) ml， P＜0.01〕. There was no statistical difference of the number of cleared lymph nodes between two groups 〔(11.8±1.5) pieces vs. (13.3±1.7) pieces， Pgt;0.05〕. The length of distal incision margin of rectal anterior resection in laparoscopic group was obviously longer than that in open group 〔(3.1±0.4) cm vs. (2.6±0.3) cm， P＜0.01〕. The gastrointestinal and urinary function of laparoscopic group recovered more quickly than those in open group 〔(2.3±0.7) d vs. (3.6±0.9) d for intake of liquid diet， P＜0.05； (3.5±1.1) d vs. (4.7±1.2) d for intake of solid diet， P＜0.05； (2.3±0.4) d vs. (4.4±1.2) d for duration of urethral catheterization， P＜0.01， respectively〕. The length of hospital stay in laparoscopic group was shorter than that in open group 〔(8.5±0.7) d vs. (12.8±0.9) d， P＜0.01〕. But the cost of hospitalization in laparoscopic group was higher than that in open group 〔(3.14±0.25)×104 yuan vs. (2.02±0.75)×104 yuan， P＜0.05〕. There was no statistical difference of the three-year survival rate between two groups (89.5% vs. 89.1%， Pgt;0.05). Conclusion Laparoscopic radical rectectomy and colectomy for colorectal cancer is feasible and safe with minimal invasiveness.