Objective To investigate the inhibitory effects of RNA interference (RNAi) expression vector on the expression of survivin in pancreatic cancer cell PANC-1. Methods The protein and mRNA expressions of survivin were examined with immunofluorescence and RT-PCR. The survivin gene was cloned into the T-vector and sequenced. The RNAi expression vectors targeting survivin, named si-svv-1 and si-svv-2 respectively according to whether they harbored a mutation or no mutation, were constructed and transfected into PANC-1 cells with liposome. The expression of survivin mRNA was detected with RT-PCR. Apoptosis of PANC-1 cells was analyzed with DNA ladder and FACS. Results There was a high degree expression of survivin in PANC-1 cells. The expression of survivin was not inhibited by RNAi expression vectors si-svv-1, but inhibited about (72.43±8.04)% by si-svv-2 and the apoptosis rate of PANC-1 cells increased to （12.36±1.44）% after 72 h. Conclusion The RNAi expression vector can effectively inhibit the expression of survivin in pancreatic cancer cell PANC-1 cells and induce the apoptosis in PANC-1 cells.
ObjectiveTo investigate the relationship between insulin-like growth factor binding protein (IGFBP) gene with pancreatic cancer.
MethodsThe relevant literatures at home and abroad in recent years were reviewed. From the pancreatic cancer related genes, IGFBP related tumors and the correlation between IGFBP and pancreatic cancer research and other aspects of the previous research results were summaried.
ResultsMost of the studies suggested that IGFBP could inhibit the function of tumor cells through the IGF dependent pathway, but the deletion or mutation of IGFBP gene and its regulation mechanism are still unclear.
ConclusionIGFBP is closely related to the tumor, but its specific effects and mechanism of pancreatic cancer has not been settled. In order to affect the degree of cell differentiation, regulation of tumor growth and metastasis probability through the change of endogenous IGFBP gene level, the further studie is needed.
Objective To investigate the effect of common iliac vein allograft replacing the portal vein-superior mesenteric vein transition area invaded by pancreatic cancer. Methods The clinical data of a patient with pancreatic cancer admitted to the Beijing Tsinghua Changgung Hospital in December 2021 who underwent pancreaticoduodenectomy combined with common iliac vein allograft replacing the junction of portal vein, superior mesenteric vein and splenic vein were analyzed retrospectively. The patient was a 77-year-old man who complained of “epigastric pain for 1 month and pancreatic mass was found for 1 week”. After admission, the patient was diagnosed with pancreatic cancer through inspection, and then the surgery was required. Preoperative examination and intraoperative exploration confirmed that the junction of portal vein, superior mesenteric vein, and spleen vein was invaded by tumor. In addition, the length of the invaded vessels measured by preoperative 3D reconstruction image was 5.5 cm, and the distance between the broken end of portal vein and the broken end of superior mesenteric vein measured was 4.5 cm during the operation. After tumor and vessels were resected, vascular anastomosis could not be performed directly. After accurate evaluation, pancreaticoduodenectomy combined with common iliac vein allograft replacing the junction of portal vein, superior mesenteric vein and splenic vein was performed. The operative time was 11 h, and the intraoperative blood loss was 400 mL. After the operation, the routine treatment was performed in ICU and was transferred to the general ward on the 7th day. Postoperative laboratory tests were performed to monitor liver function changes routinely, and imaging examination were was performed to monitor portal venous system blood flow. Results Postoperative complications such as biliary fistula, pancreatic fistula, hemorrhage, infection and thrombosis were not occurred. Postoperative pathological diagnosis: pancreatic ductal adenocarcinoma, medium-low differentiation. Enhanced CT reexamination on the 2nd and 13th day after the operation showed that the blood flow at the junction of portal vein, superior mesenteric vein and splenic vein of the common iliac vein allograft was unobstructed, and there was no stenosis or thrombosis at each anastomosis. Conclusions The application of common iliac vein allograft replacing the portal vein-superior mesenteric vein transition area invaded by pancreatic cancer is safe and feasible. The short-term efficacy is satisfactory, and long-term prognosis remains to be further observed.
ObjectiveTo systematically review the expression of E-cadherin protein and the risk of pancreatic cancer. MethodsWe searched PubMed, EMbase, The Cochrane Library, CNKI, VIP, CBM and WanFang Data from inception to October 2016 to collect case-control studies about the correlation between E-cadherin protein expression and the risk of pancreatic cancer. Two reviewers independently screened the literature, extracted data and assessed the risk of bias of included studies. Then meta-analysis was performed using RevMan 5.2 software and Stata 12.0 software. ResultsSeventeen studies (986 cases in pancreatic cancer group and 433 cases in normal pancreatic tissue group) were finally included. The results of meta-analysis showed that: the expression of E-cadherin protein in the pancreatic cancer group was lower than normal tissue group (OR=0.04, 95%CI 0.01 to 0.23, P=0.000 2), poor differentiation group was lower than high or middle differentiation group (OR=0.44, 95%CI 0.26 to 0.76, P=0.003), lymph node metastasis group was lower than without lymph node metastasis group (OR=0.50, 95% CI 0.31 to 0.81, P=0.005), and the difference was statistically significant. However, there was no significant difference between the clinical stageⅠ-Ⅱ group and Ⅲ-Ⅳ group (OR=0.63, 95%CI 0.25 to 1.59, P=0.33), pancreatic head cancer group and pancreatic body and tail cancer group (OR=1.22, 95%CI 0.72 to 2.07, P=0.46), pancreatic cancer with nerve invasion group and without nerve invasion group (OR=1.45, 95%CI 0.81 to 2.62, P=0.21), pancreatic cancer with vascular invasion group and without vascular invasion group (OR=0.55, 95%CI 0.13 to 2.22, P=0.40). ConclusionLower expression of E-cadherin protein is significantly associated with the risk of pancreatic cancer. Due to the limited quality and quanity of includied studies, the above conclusion should be approved by more studies.
ObjectiveTo systematically evaluate the potential value of C-reactive protein to albumin ratio (CAR) as an indicator of prognosis and survival in patients with pancreatic cancer. MethodsThe literatures were searched comprehensively in the PubMed, Embase, Web of Science, Cochrane Library, CBM, Wanfang, CNKI, and CQVIP databases from the establishment of the databases to May 20, 2021. The combined hazard ratio (HR) and 95% confidence interval (95%CI) were used to evaluate the correlation between the CAR and the overall survival (OS), progression-free survival (PFS), or disease-free survival (DFS) in the patients with pancreatic cancer. The Newcastle-Ottawa scale (NOS) was used to evaluate the quality of the non-randomized controlled studies, and the Stata SE 15.0 software was used for meta-analysis. ResultsA total of 2 985 patients with pancreatic cancer were included in this meta-analysis of 15 studies. The results of meta-analysis showed that the higher CAR value, the shorter OS [effect size (ES)=0.60, 95%CI (0.50, 0.69), Z=12.04, P<0.001], DFS [ES=0.63, 95%CI (0.47, 0.78), Z=3.61, P<0.001], and PFS [ES=0.41, 95%CI (0.19, 0.63), Z=7.91, P<0.001] in the patients with pancreatic cancer. The results of subgroup analysis of OS according to different countries, sample size, mean age, follow-up time, CAR cut-off value, and NOS score showed that the higher CAR value was related to the shorter OS (P<0.05). The result of linear regression analysis showed that there was no correlation between the CAR cut-off value and lnHR of OS (r2=0.947, P=0.455). Conclusion From results of this study, CAR is closely related to OS of patients, and it is expected to be used as a new reference index for monitoring and judging prognosis of patients with pancreatic cancer.
Objective To investigate the expressions of CXCR4 and β-catenin in pancreatic cancer, explore the relationship between them, and explore the possible biomarkers about invasion and metastasis of pancreatic cancer. Methods Forty-eight samples of pancreatic cancer and 20 samples of normal pancreas tissues were selected. The expressions of CXCR4 and β-catenin were examined by the immunohistological technique. Spearman, Chi-square， and rank test were used to analyze the relation between the protein expressions and clinical characteristics. Survival analysis was evaluated by Kaplan-Meier product limit method and Log-rank test. Variables were evaluated by Cox proportional hazards analysis. The size of test was 0.05. Results The positive expression rates of CXCR4 and β-catenin in pancreatic cancer tissues were 85.4% (41/48) and 75.0% (36/48), respectively. Co-expression rate of CXCR4 and β-catenin was 70.8% (34/48). There were significant differences between various CXCR4 staining and lymph node metastasis and TNM stage (P=0.012, 0.005, respectively). β-catenin positive expression was associated with lymph node metastasis (P=0.047). However, abnormal β-catenin positive expression could not determine the clinical survival. Kaplan-Meier estimated curves suggested that clinical prognosis was poor for patients with CXCR4 expression. Multivariate analysis showed that CXCR4, late TNM stage, and lymph node metastasis were independent prognostic factors for pancreatic cancer. Conclusions Both CXCR4 and β-catenin abnormally express in pancreatic cancer. CXCR4 may be an important marker for pancreatic cancer progression.
Objective
To systematically review the diagnostic value of combined detection of K-ras gene mutation in peripheral blood and serum tumor markers for pancreatic cancer.
Methods
Databases including PubMed, The Cochrane Library (Issue 3, 2016), Elsevier, BMJ, CBM, CNKI and WanFang Data were searched from 2000 to March 2016 to collect diagnostic tests about the diagnostic value of combined detection of K-ras gene mutation in peripheral blood and serum tumor markers in pancreatic cancer. Two reviewers independently screened literature, extracted data and assessed the methodological quality of included studies. Then, meta-analysis was performed using RevMan 5.3 software.
Results
A total of 23 studies involving 2?071 patients were included. The results of meta-analysis showed that the sensitivity (SE) and specificity (SP) of K-ras gene mutation in peripheral blood were 65% and 92% respectively in the diagnosis for pancreatic cancer. The results of the detection of tumor marker CA19-9 were 78% and 81% respectively. The SE and SP indexes in the parallel and serial combinations of CA19-9 together with CA242 were 85%, 72%, 70% and 83% respectively. And the SE, SP indexes in the parallel and serial combinations of K-ras gene mutation combined detection with CA19-9 were 90%, 63%, 47% and 96%. The positive likelihood ratio of the parallel combination of K-ras gene mutation in peripheral blood and CA19-9 (+LR=10.89) was higher than the other three detection methods, while the negative likelihood ratio of the serial combination of K-ras gene mutation in peripheral blood and CA19-9 (-LR=0.15) was lower than the other three detection methods, which indicated that the combined detection of K-ras gene mutation in peripheral blood and and CA19-9 had a better diagnostic performance than the single dectection of K-ras gene mutation or CA19-9 or the combined detection of CA19-9 and CA242 respectively. Comparing the area under curve (AUC) of SROC curve of the two combined diagnoses, the results showed that the diagnostic value of the parallel combination of K-ras gene mutation in peripheral blood and CA19-9 (AUC=0.87) was higher than that of the parallel combination of serum tumor markers CA19-9 and CA242.
Conclusion
Current evidence indicates that the combined detection of K-ras gene mutation and and tumor marker CA19-9 levels in peripheral blood can improve the diagnostic accuracy for pancreatic cancer. Due to the limited quantity and quality of included studies, above conclusions need to be verified by conducting more high quality studies.
Objective To investigate the inhibitory effect of survivin antisense oligonucleotides (ASODN) on proliferation of pancreatic cancer cells PANC-1. Methods The ASODN and sense oligodeoxynucleotides (SODN) were complementary to survivin sequences. FAM-marked ASODN was transfected into PANC-1 cells mediated by positive ion liposome as ASODN group. Blank control group (normal cells), negative control group (normal medium)， and SODN group were established for comparison. The transfection efficiency was detected by flow cytometry (FCM) after transfection; MTT assay was used to detect cytotoxicity; Cell morphological changes were examined by transmission electron microscopy; The cell cycle and apoptotic rate were analyzed by FCM; Immunohistochemical staining techniques were used, and the expressions of survivin were observed under light microscopy, examined and analysed by computer image. Results ①The transfection efficiency was 31.9%, 37.4%, 41.4%, 52.6%, 24.2%, 11.4%, 16.1%, and 15.5% when the transfecting concentration of ASODN was 50, 100, 150, 200, 250, 400, 600, and 800 nmol/L, respectively; The transfection efficiency was 12.0%, 50.8%, and 11.2% when the inoculated cells was 2×104/well, 2×105/well, and 2×106/well, respectively; The transfection efficiency was 58.8%, 34.0%, and 23.6% when 2 μl, 3 μl, and 4 μl liposome was used during transfection, respectively. ②Cell gap was oversize, morphous was round, adherent cells were less after transfection under fluorescence microscope. ③The inhibition rate in the ASODN group was higher than that in each control group (Plt;0.05) on 24, 36, 48 h after treating by survivin ASODN, which increased as time prolonged (Plt;0.05). ④The apoptosis showed a ladder-shaped line in the ASODN group. ⑤Apoptotic morphology was demonstrated in the ASODN group, such as apoptotic cells with nuclear chromatin highly concentrated, crescent nuclear staining aggregated by the side nuclear membrane, nucleolus disappeared by AO and EB stains. ⑥The apoptotic rate 〔（38.1±3.4）%〕 in the ASODN group was higher than that in the SODN group 〔（4.16±1.7）%〕, Plt;0.05. ⑦G2/M cell cycle arrested in the ASODN group. ⑧After transfection, the expression of survivin protein in the ASODN group was significantly lower than that of each control group (Plt;0.05). Conclusions The optimal transfection conditions are as following: the cell count of 2×105/well, concentration of ASODN 200 nmol/L, and cationic liposome oligofectamine 2 μl, respectively. Survivin ASODN can inhibit the proliferation of pancreatic cancer cells and induce their apoptosis.
Objective To investigate the effect of angiostatin gene combined with somastatin on inhibiting proliferation of human pancreatic cancer cell BXPC-3 and endothelial cell of vascular ECV-304 and on inducing their apoptosis in vitro. Methods The pcDNA3/angio was transfected BXPC-3 by liposome-mediated gene transfer method. RT-PCR and Western blot were used to detect the expression of angiostatin gene. In vitro, MTT and flow cytometry (FCM) were used to detect whether angiostatin gene combined with somastatin could effect the growth inhibition of BXPC-3 and ECV-304 cells. Results Angiostatin was expressed and secreted by transfected BXPC-3. The growth of BXPC-3 was inhibited by certain concentration of somatostatin (≥10 μg/ml, P＜0.01）, which was dependent on the dose of somatostatin in a concentration extent; Simultaneity apoptosis was induced （P＜0.01）. But the growth of ECV-304 was not inhibited with somatostation （Pgt;0.05）. Angiostatin could inhibit the growth of ECV-304 and induced apoptosis （P＜0.01）, but it had no effect on the growth of BXPC-3 （Pgt;0.05）. Angiostation gene combined with somatostation could inhibit the growth both of BXPC-3 and ECV-304 （P＜0.01）, and induce apoptosis of them （P＜0.01）; but the effect couldn’t be additived. Conclusions ①Somatostatin directly inhibits the proliferation of human pancreatic cancer cells and induces apoptosis, but it doesn’t directly inhibit angiogenesiso of human pancreatic cancer. ②Angiostatin specially inhibits the proliferation of endothelial cell of vascular and induces apoptosis. Angiostatin could inhibit angiogenesis of human pancreatic cancer to induce necrosis of cancer cell.
ObjectiveTo explore the values of CA19-9, CA242, CEA, and CA125 single or combined detection on clinical diagnosis and prognosis for patients with pancreatic cancer. MethodsSerum tumor markers CA199, CA242, CEA, and CA125 of 63 patients with pancreatic cancer, 33 patients with cancer of bile duct, and 27 patients with benign pancreatic disease were detected, and those patients were followed up after operation. ResultsThe levels of CA19-9, CA242, CEA, and CA125 in patients with pancreatic cancer were significantly higher than those in patients with benign pancreatic disease and cancer of bile duct (Plt;0.05). The sensitivity of CA19-9 alone was the highest in the four tumor markers for the patients with pancreatic cancer 〔79.4% (50/63)〕, but the specificity (61.9%) was lower than that of CA242 (83.3%) and CEA (80.0%). The specificity of combined detection of CA199+CA242+CEA was the highest 〔93.3% (56/60)〕. The level of CA19-9 in carcinoma of body/tail of pancreas was significantly higher than that of carcinoma of pancreas head or whole pancreas (Plt;0.05). The serum levels of CA19-9 and CA242 in patients with stage Ⅳ were significantly higher than those in stage Ⅰ or Ⅱ/Ⅲ (Plt;0.05). Fifteen patients were lost to follow up, 48 patients were followed up 2－12 months with an average 6 months. The levels of CA242 and CA199 in patients with pancreatic cancer on 0.5 month and 3 months after operation were lower than those before operation (Plt;0.05). ConclusionsSingle detection of CA19-9 can improve the diagnostic sensitivity, and combined detection of tumor markers CA199+CA242+CEA can improve the diagnostic specificity. CA19-9 or CA242 is a valuable marker for evaluating treatment effects and estimating prognosis.
