A Retrospective Survival Analysis Of Post-interventional-therapy Patients With Unresectable Pancreatic Carcinomas And An Experimental Research On MicroRNAs Associated With Gemcitabine-resistance

Objective:To evaluate important factors for survival time and research effective interventional therapy programs by a retrospective survival analysis of post-interventional-therapy patients with unresectable pancreatic carcinomas.Materials and methods:Clinical records of159patients with unresectable pancreatic carcinomas were collected and retrospectively analysis, univariate and Cox Model multivariate statistical analysis were applied to evaluate factors which influence survival time and effective programs for patients with unresectable pancreatic carcinomas.Results:The total following-up median survival time (MST) of included159patients was10.32months and the MST of post-interventional-therapy was8.11months. The cumulative survival rates of1-year,2-year,3-year, and5-year were27.0%,11.0%,2.0%and0.6%. The averaged times of interventional therapy which could alleviate symptoms were2.6, judged by clinical benefit rate (CBR). Univariate analysis showed that the important factors for post-interventional-therapy MST were blood CA199value before treatment, blood CA199change after first treatment, KPS scores before treatment, symptoms before first treatment, liver function grade before treatment, operation history of primary site, interventional therapy times. There were no significant difference in age, sex, blood haemoglobin(Hb),tumor stage(stage II-III compare to stage IV), blood CEA before treatment, blood CEA change after first treatment, GEMOX (gemcitabine plus oxaliplatin)or not, transcatheter arterial infusion(TAI) or transcatheter arterial chemoembolization(TACE), Diabetes Mellitus(DM) or not, pancreatic head or not(primary cancer site),pancreatic ductal adenocarcinoma or not. Multivariate analysis indicated that the independent factors for survival time were KPS scores before treatment, liver function grade before treatment and operation history of primary site.Conclusion:Interventional therapies including TAI and TACE could offer some clinical benefits:improvement of symptoms, prolongation of MST. Post-interventional-therapy survival time was associated with body status, symptoms and operation history of primary site. Early diagnosis and early treatment as well as research of reducing drug-resistance of pancreatic cancer are our urgent tasks in the future. Objective:To determine the safety and efficiency of21G FNAC compared to18G Tru-cut core needle (TCN) in liver tumor biopsies, and the safety and efficiency of21G FNAC biopsy applied in pancreatic tumors.Patients and methods:1.94patients who were diagnosed as unresectable malignant liver tumors and were going to undergo transarterial chemoembolization (TACE) were included. Before treatments, percutaneous liver biopsies were arranged. Data was collected and all patients were divided into2groups:group18G TCN(every biopsy was finished with a18G True-cut core needle) and group21G FNAC. All biopsies were guided by B ultrasonic (BUS).In group FNAC, a21G fine needle was used to puncture into the targeted mass, then the stylet was pulled out. The core needle was pushed and pulled for several times in the body of the targeted mass so that tissues of the tumor can be cut off and filled into the needle. At the same time a negative pressure was maintained by a syringe connected to the needle. All tissues and absorbed fluids were undergone pathologic and cytological examination. After the biopsy, an arterial angiography was carried out to judge if bleeding in needle tracks existed. TACE was performed according to the situation of the patient. The total positive rate (TPR) was obtained by counting positive rate of pathologic and cytological examination. The TPR and security of both groups were statistically analysed and compared.2.21G FNAC was applied in18patients with pancreatic tumors and the safety and TPR was analysed. Results:All biopsies of selected patients included liver and pancreatic tumors were successfully fulfilled. The TPR of18G TCN is89.1%(41/46).The incidence rate of subcapsular hemorrhage of liver punctured track (according to arterial angiography) of group18G TCN was6.5%(3/46). Incidence rate of pain(caused by biopsy) was78.3%(36/46).There was8.7%(4/46) incidence rate of arteriovenous shunt(A-V shunt) in group18G TCN. The TPR of group21G FNAC was81.3%(39/48). There was no statistically significant difference of TPRs between the two groups of liver biopsy (P>0.05). It showed that there was statistically significant difference between the two groups in tissue length. There were82.6%(38/46) patients whose samples’length>0.5cm in group18G TCN while52.1%(25/48) in group21G FANC. Enough tissues could be obtained for immunohistochemical exams in87.0%(40/46) patients of group18G TCN and75.0%(36/48) of group21G FNAC. Incidence rate of pain(caused by biopsy) was22.9%(11/48) in group18G TCN. There was statistically significant difference of pain between the2groups of liver biopsy. All patients felt slight pain(VAS<3) at puncture points and recovered2-3days later without any pain-killer. No patients suffered from pneumothorax, pleural effusions, and peritonitis. There were no patients evolving severe complications such as massive bleeding and exsanguine shock.No tumor implantation was found in needle tracks after3months follow-up. The TPR of18patients with pancreatic tumors was61.1%(11/18).Incidence rate of pain was27%(5/18)and incidence of pancreatitis was0.56%(1/18).There were no other complications occurred.Conclusion:TPR was not different between the21G FNAC and18G True-cut biopsy procedures, but the safety of21G FNAC may be superior to that of18G Tru-cut biopsy. Liver tumor biopsy tissue obtained using either of these two procedures can result in a pathologic diagnosis, according to patient’s situation.21G FNAC biopsy was suit for pancreatic tumor for its safety and efficiency. Objectives:To search miRNAs associated with gemcitabine-resistance formed by in vitro simulation of pancreatic cancer cells exposed to gemcitabine and explore the regulation of gene expression of interesting miRNAs, as well as the guiding significance of clinical medicine.Materials and Methods:BxPC-3pancreatic cancer cell lines were cultivated into gemcitabine-resistant strains with increasing concentrations and induction of short-term role. Logarithmic growth phase cells were added into gemcitabine solution from low concentrations of2ng/ml increase to30ng/ml for48hours every kind of concentration. The sensitive dead cells were cleaned by changing new culture fluid.Live cells were kept and continuously incubated. Gemcitabine-resistant cell lines were induced combined with continuously drug screening. BxPC-3pancreatic cancer cells and gemcitabine-resistant cell lines were undergone miRNA microarray detection and compared to find down-regulated miRNAs.The major regulatory genes of these miRNAs were analyzed. Real-time PCR and Western Bolt were performance to detect the expression of these genes in drug-resisitance formation,and microarray results were validated.Results:Gemcitabine-resistant cell lines of BxPC-3were successfully established.By chip detection and analysis, comparing to the parental cells,we found that there were11miRNAs which expressed significantly up-regulated in cells resistant strains and23significantly downward. Network-control analysis results showed that the degree of has-mir-20a in the regulatory network was the highest one and the degree of its target genes regulated ABCG2was the highest too. Real-time PCR results showed that, compared to the parental cells, has-mir-20a was downregulated in gemcitabine-resistant strains.lt validated the results of microarray. ABCG2expression was detected to be significantly upregulated by Western bolt test.Conclusion:Has-mir-20a may play an important role in formation the gemcitabine-resistance. The down-regulated expression of has-mir-20a led to the up-regulated expression of ABCG2, which is closely related to drug-resistance formation and pancreatic cancer.