Lineage plasticity-mediated therapy resistance in prostate cancer / 亚洲男科学杂志(英文版)

Therapy resistance is a significant challenge for prostate cancer treatment in clinic. Although targeted therapies such as androgen deprivation and androgen receptor (AR) inhibition are effective initially, tumor cells eventually evade these strategies through multiple mechanisms. Lineage reprogramming in response to hormone therapy represents a key mechanism that is increasingly observed. The studies in this area have revealed specific combinations of alterations present in adenocarcinomas that provide cells with the ability to transdifferentiate and perpetuate AR-independent tumor growth after androgen-based therapies. Interestingly, several master regulators have been identified that drive plasticity, some of which also play key roles during development and differentiation of the cell lineages in the normal prostate. Thus, further study of each AR-independent tumor type and understanding underlying mechanisms are warranted to develop combinational therapies that combat lineage plasticity in prostate cancer.

A simplified method for isolating murine primary pancreatic acinar cells / 中华胰腺病杂志

Objective To explore a simplified and economical method to isolate the murine primary pancreatic acinar cells.Methods The collagenase and trypsin inhibitor dissolving in DMEM solution were used to digest the murine pancreas,and 4％ BSA dissolving in DMEM solution was used to purify and isolate primary pancreatic acinar cells from pancreas.CCK-8 method was applied to check the ability of pancreatic acinar cells to secret amylase.Results After digestion,shaking in the water bath,resuspension,filtration and precipitation,murine primary pancreatic acinar cells could be obtained within 2 hours.Pancreatic acinar cells in good conditions appeared in clusters,and their basolateral domains were round and devoid of blebs,and the cytoplasm appeared clear.Their apical domain were surrounded by hundreds of zymogen granules which looked darker.The nucleus was located in the basal area of the vesicular region.The basal level of amylase release as a percent of total release from pancreatic acinar cells was around 2.5％ in CCK8-unstimulated group.This rate started to increase after CCK-8 stimulation and reached its peak [(12.83 ± 1.04) ％] at a concentration of 50 pmol/L of CCK-8,but the ratio of the amylase level secreted by the pancreatic acinar cells to the total amylase level displayed a decreasing trend with the increase of CCK-8 concentration.Conclusions This optimized method had the advantage of being fast and simple,low technical difficulty and good repetition.It was a new simplified and cheap method for isolating murine pancreatic acinar cells.

Effects of ApoC3 on the severity of acute pancreatitis in mice / 中华胰腺病杂志

Objective To explore the effects of ApoC3 gene on the severity of hypertriglyceridemiainduced acute pancreatitis (AP).Methods ApoC3 transgenetic mice and C57BL/6J mice AP model was induced by cerulein intraperitoneal injection,and ApoC3 transgenetic mice and C57BL/6J mice injected by normal saline solution in equal volume served as control group.Serum triglyceride and cholesterol were detected,and the pathological changes of the pancreas were observed.RT PCR method was used to examine the changes of the inflammatory factor including IL-1β,IL-6,α-SMA and TNF-α mRNA levels,which reflected the severity of the inflammation.Results Serum triglyceride and cholesterol were higher in ApoC3 transgenetic mice than in C57BL/6J mice [(3.434 ± 0.931) mmol/L vs (0.766 ± 0.120) mmol/L,(2.553 ±0.178) mmol/L vs (1.996 ± 0.080) mmol/L],and the differences were statistically different (P ＜ 0.05).The pathological changes of the pancreas were more severe in ApoC3 transgenetic AP mice than in C57BL/6J AP mice,and the IL-1β,IL-6 and α-SMA mRNA levels in the pancreatic tissue were obviously higher in ApoC3 transgenetic AP mice than in C57BL/6J mice (1.72 ± 0.07vs 0.78 ± 0.09,1.58 ± 0.09vs 0.87 ±0.04,0.83 ± 0.05vs 0.44 ± 0.04),and the differences were statistically significant (P ＜ 0.05),while there was no statistical difference on TNF-αmRNA level (0.70 ± 0.09vs 0.65 ± 0.08,P ＞ 0.05).Conclusions ApoC3 gene could aggravate the severity of the inflammation in hypertriglyceridemia-induced AP.

The expression of HMGB in rats with acute necrotizing pancreatitis / 中华胰腺病杂志

Objectives To detect the expression of serum high mobility group box-1 (HMGB1) and explore its changes in rats with acute necrotizing pancreatitis (ANP).Methods Intraperitoneal injection of 20％ L-arginine in the dosage of 250 mg/100 g twice every 1 hour was used to establish ANP rat model.Intraperitoneal injection of normal saline solution in equal volume was performed in control rats.Rats were sacrificed at 6 h,18 h,24 h,36 h,48 h,72 h and 96 h after injection.Blood samples were collected to detect serum amylase and HMGB1 level.Pancreatic tissue was collected for pathological examination.Realtime PCR was applied to detect the mRNA expression of HMGB1 in pancreatic tissue.Werstem blot was used to determine HMGB1 protein expression in pancreatic tissue.Results Serum amylase level began to increase at 6 h after modeling,reached the peak at 18 h [(5 070 ± 603) U/L] and returned to normal level after 48 h.Serum amylase activity at 6 h and 18 h in ANP group was much higher than that in control group (1 844 ± 181)U/L(P＜0.05).The expression of HMGB1 began to increase at 6 h,reached to the peak at 36 h [(288.5 ±42.1)μg/L],and then decreased gradually.HMGB1 expressions at each time point in ANP group were significantly higher than those in control group (31.6 ± 10.1) μg/L],and the differences were statistically significant (all P ＜ 0.05).Pathological scores in pancreatic tissues in ANP group were higher than those in control group 0.38 ± 0.52,and the differences were statistically significant (P ＜ 0.05).HMGB1 mRNA expressions at t 6 h,18 h,24 h,36 h,48 h,72 h and 96 h in ANP group were 1.23 ±0.25,2.60 ± 0.46,3.23 ± 0.34,4.77 ± 0.66,2.88 ± 0.56,2.05 ± 0.20,1.33 ± 0.28,which were significantly higher than those in control group 0.44 ± 0.09,and the relative expression of HMGB1 in ANP group at 36 h was significantly higher than those at other time points (all P ＜ 0.05).HMGB1 protein expression in pancreatic tissue in ANP group at 6 h,18 h,36 h,72 h were 1.14 ±0.02,1.15 ±0.01,1.22 ±0.01,1.22 ±0.04,which obviously higher than those in control group(1.0),and HMGB1 expression in ANP group at 36 h was higher than those at other time points (all P ＜ 0.05).Conclusions HMGB1 may participate in systematic inflammation as one of the late inflammatory mediators during ANP.

Clinical significance of the methylation in TPAP promoter in pancreatic cancer / 中华胰腺病杂志

Objective To investigate functional role and clinical significance of the methylation in the promoter of TPAP gene in the development and progression of pancreatic cancer.Methods Surgically resected specimen from 68 patients who were pathologically diagnosed as pancreatic cancer in Changhai Hospital from July 2006 to August 2009 were collected.The methylation in the promoter of TPAP gene in tumor and nontumor adjacent tissue was detected by methylation specific PCR.Results The methylation rate of tumor and non-tumor adjacent tissue was (0.214 ± 0.057) ％ and (0.084 ± 0.096) ％,respectively,and pancreatic cancer tissue had significantly higher methylation rate than the adjacent tissue.Hypermethylation of TPAP gene was not correlated with age,gender,tumor differentiation,lymphatic metastasis,serum CEA and CA19-9,but was positively correlated with distant metastasis.Conclusions Hypermethylation in the promoter of TPAP gene may participate in the invasion and metastasis of pancreatic cancer and the hypermethylation of the promoter is closely associated with the tumorigenesis and development of pancreatic cancer.

Significance of quantitatively detecting K-ras codon 12 and 13 mutations in the tissues of pancreatic cancer and related pancreatic diseases / 中华胰腺病杂志

ObjectiveTo investigate the clinical significance of quantitative detection of K-ras codon 12 and 13 mutations in the tissues of pancreatic cancer and related pancreatic diseaaes. Methods One hundred and thirty samples from surgically removed pancreatic tissue with a conclusive pathological diagnosis (105 cases of pancreatic ductal adenocarcinoma,8 cases of pancreatic adenosquamous carcinoma of the pancreas,2 cases of pancreatic mucinous adenocarcinoma,3 cases of pancreatic endocrine carcinoma,6 cases of duodenal and papillary adenocarcinoma and 6 cases of benign pancreatic diseases ) were collected.Quantitative detection of K-ras codon 12 and 13 mutations was performed by the method of peptide nucleic acidmediated PCR clamping with two different fluorescence labeled probes.Mutation number ＞ 100 copies was used as the criteria to calculate the positive mutation rate.ResultsThe median and quartile of K-ras codon 12mutations of pancreatic ductal adenocarcinoma,adenosquamous carcinoma of the pancreas,pancreatic mucinous adenocarcinoma,pancreatic endocrine carcinoma,duodenal and papillary adenocarcinoma and benign pancreatic diseases were 4062 (495,10800),238 (45,8420),15 (9,21),3 (3,16),2283 (73,5037)and 21(8,56),and the positive mutation rates were 84.8％ (89/105),50.0％ (4/8),0,0,66.7％ (4/6)and 16.7％ (1/6).The quantity of K-ras codon 12 mutation in pancreatic ductal adenocarcinoma was not statistically different from those of adenosquamous carcinoma,duodenal and papillary adenocarcinoma,but it was significantly higher than those in pancreatic mucinous adenocarcinoma,pancreatic endocrine carcinoma,and benign pancreatic diseases (P ＜0.05).The area under ROC of K-ras codon 12 mutation in pancreatic ductal adenocarcinoma was 0.727.The sensitivity and specificity of the K-ras codon 12 mutation for the diagnosis of pancreatic ductal adenocarcinoma were 84.8％,64.0％,respectively.The quantity of K-ras codon 12 was associated with survival of patients with pancreatic ductal adenocarcinoma.The quantity of K-ras codon 13 mutations and the positive mutation rates in pancreatic ductal adenocarcinoma was not statistically different from other pancreatic diseases.ConclusionsThe quantity of K-ras codon 12 mutation has good differential diagnostic and prognostic prediction value for pancreatic ductal adenocarcinoma.

Quantitative detection of mutation of K-ras gene at codon 12 / 中华胰腺病杂志

Objective To quantitatively analyze the K-ras gene mutation at codon 12 in pancreatic cancer tissues and the relationship between K-ras gene mutation and clinicopathological parameters. Methods Quantitative detection of K-ras gene at codon 12 in 93 pairs of pancreatic cancer and adjacent tissues were performed by using PNA-mediated PCR clamping with two different fluorescence labeled probes. The quantity of mutation was expressed by percentage of mutation. The percentage of K-ras gene mutation = the copy of K-ras mutation/(copy of wild type K-ras + copy of K-ras mutation) × 100%. Results The percentage of mutation of K-ras gene at codon 12 in pancreatic cancer and adjacent tissues were 83.9% and 65.6%, and the difference was statistically significant(P ＜ 0. 05); and the quantity of mutation were (13.385 ± 1. 745) % and (2. 246 ±0. 728) %, and the difference was also statistically significant(P ＜ 0. 05). The quantity of mutation of K-ras gene at codon 12 was not associated with clinicopathological parameters. Conclusions The percentage of K-ras gene mutation, as well as the quantity of K-ras gene mutation was different in pancreatic carcinoma and adjacent tissues.

Expression of microRNAs in fecal of patients for pancreatic cancer screening and detection / 中华胰腺病杂志

Objective To detect the microRNAs in fecal with patients of pancreatic cancer, and evaluate its diagnostic value. Methods Stool samples were collected from three group persons including 29 pancreatic cancer, 22 chronic pancreatitis and 13 normal controls. The total fecal microRNAs were extracted.The quantity of miR-16, miR-21, miR-155, miR-181a, miR-181b, miR-196a, and miR-210 were detected by using real-time PCR, and miR-16 was used as reference gene. ROC AUC was used to evaluate the diagnostic value for pancreatic cancer. Results MicroRNAs were efficiently obtained from stools, and independent experiments showed high reproducibility for microRNAs extraction and detection. The expression of miR-181b,miR-196a, miR-210 in fecal was 2.22 ±0.64,2.78 ±0.14, 5.55 ±0.38 in pancreatic cancer; 1.42 ±0.39,3.88 ± 0.85,5.39 ± 0.69 in chronic pancreatitis; 0.32 ± 0.40, 1.14 ± 0.98,4.23 ± 0. 99 in normal controls;the three microRNA expressions in pancreatic cancer were group and CP group significantly higher than those in normal controls ( P ＜ 0.05 ). But there was no significant difference between pancreatic cancer group and chronic pancreatitis group. AUC of pancreatic cancer / normal controls miR 18lb was 0.745(95% CI 0. 597-0.894), the sentivity, specificity for pancreatic cancer was 84.6% and 51.7%. AUC of miR-210 was 0. 772(95% CI0.629-0.914), the sentivity, specificity for pancreatic cancer was 84.6% and 65.5%, and the difference was statistically significant (P ＜0.05). miR-196a was no significant for the diagnosis of pancreatic cancer, but the expression of miR-196a was correlated with the tumor size (r = 0.516, P = 0.041 ).Conclusions The extraction and detection of the fecal microRNAs were non-invasive and reproducible. The expression of miR-181b and miR-210 was increased in stool of patients with pancreatic cancer, and may be potential biomarker for pancreatic cancer.

Methylation of PCDH8 in pancreatic carcinoma cell lines / 中华胰腺病杂志

Objective To investigate the methylation status of PCDH8 gene in pancreatic carcinoma.Methods Methylation of PCDH8 gene in 2 samples of normal pancreatic tissues and 6 pancreatic carcinoma cell lines (PANC1, ASPC1, BxPC3, CFPAC, PaTu8988 and SW1990) was detected by the methylationspecific PCR (MSP) method. The expression of PCDH8 mRNA was detected with 5-Aza-2-deoxycytidine (5-Aza-dC) treatment, a kind of DNA methyltransferase (DNMT) inhibitor in 6 pancreatic carcinoma cell lines by real-time-PCR. Results The methylation of PCDH8 gene was not detected in normal tissues, while it was partially methylated in PANC1, BxPC3, CFPAC and it was totally methylated in PaTu8988, ASPC1, SW1990.PCDH8 mRNA was expressed in PANC1, SW1990, PaTu8988 and the relative quantities of mRNA expression (RQ) were 1.576 ± 0.648, 0.013 ± 0.008, 0.002 ± 0.001; PCDH8 mRNA was not expressed in BxPC3,CFPAC, ASPC1. After 5-Aza-dC treatment, PCDH8 mRNA was expressed in PANC1, ASPC1, BxPC3,CFPAC, PaTu8988, SW1990 and the relative quantities of mRNA expression all significantly increased, and they were 7. 463 ± 2.628, 10. 696 ± 1.539, 7.852 ± 2.762,421.815 ± 1.493, 118.595 ± 4.089, 6.690 ±1.884. Conclusions The methylation of PCDH8 gene may be the major mechanism of down-regulated expression of PCDH8 gene in pancreatic carcinoma.

The value of the K-ras mutations in FNA samples of pancreas on the diagnosis of pancreatic cancer / 中华胰腺病杂志

Objective To investigate the diagnostic value of the K-ras mutations in FNA samples for early detection of pancreatic cancer. Methods FNA samples of 27 patients with pancreatic cancers, 9 patients with other malignant tumors and 14 patients with non malignant pancreatic mass (NMPM) were collected. DNA was extracted, and K-ras gene was amplified through PNA-mediated PGR clamping, the products were sequenced to determine the mutation type. Results The positive rate of K-ras mutations in pancreatic cancers,other malignant tumors and NMPM were 88.9%, 44.4%, 35.7%. There was significant difference in K-ras gene mutations in FNA samples between pancreatic cancer and other malignant tumors ( P = 0. 013 ) and NMPM ( P = 0. 001 ). The sensitivity, specificity, positive predictive value, negative predictive value,accuracy of K-ras mutations in FNA samples of pancreatic cancers were 88.9%, 55.6%, 85.7%, 62.5%,80.6% when compared with other malignant tumors, and the difference between the two groups was significant (P =0. 013) ;Those were 88.9%, 64.3%, 82.8%, 75.0%, 80. 5% when compared with NMPM, and the difference between the two groups was significant ( P = 0. 001 ). When cytology of FNA samples and K-ras mutations was combined, the positive rate of pancreatic cancer was up to 96.3%. Conclusions The detection of K-ras mutations in EUS-FNA samples helped improve the positive diagnostic rate of pancreatic cancer.

Expression and clinical implications of RADIL gene in pancreatic cancer / 中华胰腺病杂志

Objective To investigate the RADIL mRNA expression in pancreatic carcinoma and to evaluate its clinical significance.Methods Fluoesecent quantitative PCR (FQ-PCR) was used to detect the RADIL mRNA expression in 40 patients with pancreatic carcinoma and adjacent tissue and in 5 healthy adult with normal pancreatic tissue and to observe its relationship with clinicopathologic parameters.Results RADIL mRNA was expressed in pancreatic carcinoma and adjacent tissue, as well as normal pancreatic tissue, and the relative expression was 2.263 ± 3.826, 5.425 ± 8.858 and 8.559 ± 4.214, respectively.There was statistically significant difference among the three groups (P ＜0.05 ).RADIL mRNA expression was closely related with the metastasis and differentiation grade ( r = -0.312 and -0.294, P ＜ 0.05 ), however, it was not significantly related to tumor site, tumor size, CA19-9, TNM staging, sex and age.Conclusions RADIL gene may have an inhibitory effect on the pancreatic cancer.

Methylation analysis of human hedgehog interacting protein gene in pancreatic juice / 中华胰腺病杂志

nd hypermethylation of HHIP was detected in pancreatic juice,which may be a useful marker in the diagnosis of PCa.