In Vitro Drug Sensitivity Test And Study On Increasing Chemosensitivity On Pancreatic Cancer

Part I Study on primary pancreatic cancer isolating,purifying and certification ObjectivePancreatic cancer has a poor prognosis, primary pancreatic cancer cell isolation could afford objective for fundament and clinical research. We will afford primary cancer cell for pancreatic cancer cell chemosensitivity testing in vitro. Methods and MaterialsPancreatic cancer tissues were broken up by collagenase, and then be filtrated. Only cancer cell would be achieved from eliminated mixed cell by differential joint time methods. At last, the cell were proved cancer cell by immunohistostaining. Result23 pancreatic tissues was digested by collagenase and 17 primary cell was successfully isolated and growth steadily. About 85±7 per cent of cells were confirmed come from epidermis tissues.Conclusion collagenase digest and differential joint time method could afford us enough cells for chemosensitivity test in vitro. Part IIAnalysis of factors on drug sensitivity test of pancreatic cancer invitroObjectIn-vitro drug sensitivity testing can be interfered by many aspects, such as concentration and touching time, cell density , mixed fibroblast cell, time of add drug to median and et al. We should control anything above in condition to analysis between inhibiting rate and drug sensitivity. Methods and materialswe regarded pancreatic cancer cell lines Panc-1 and ASPC-1 as our study object, detected Gemzar, Mitomycin and 5-FU inhibit rate in varies of concentrations and touching time using MTT assay. We Analyzed effects of other aspects on testing by orthogonal design method. ResultThe growth inhibitory of Gemzar is concentration-dependent, while 5-FU is time-dependent and mitomycin is either concentration-dependent or time-dependent. Adding drug time, Varies of cell densityand whether mixed fibroblast cell would affect drug sensitivity test(p<0.05). ConclusionDrug sensitivity test might be affected by many factors, we should control many factors in extra condition before evaluated drug sensitivity testing. Part III Verify the difference between traditional therapy and chemotherapyfiltered by drug sensitivity test in vitro ObjectPancreatic cancer is one of the most malignant cancer with poor prognosis and always resistant to chemotherapy. Gemcitabine was found to be active against pancreatic cancer. Yet Gemcitabine is always the most sensitive of all drugs? Here we filtered the most sensitive drugs from Gemcitabine,mitomycin and 5-FU, and then compared curative effects between chemotherapy filtered by drug sensitivity test in vitro and traditional therapy from animal experiment. Methods and materialsWe detected drug cytotoxin to vary of pancreatic cancer cell lines and primary cells using MTT assay and accounted the IC₅₀, then analyzed sensitivity and filtered the most sensitive drug. Established the model of transplant carcinoma in nude mice and compared effects between traditional chemotherapy and chemotherapy filtered by drug sensitivity test in vitro. ResultTwo pancreatic cancer cell lines(Panc-1 and ASPC-1) and 17 primary cell was filtered. Results showed that the most sensitive drug was 5-FU in ASPC-1 and 6 primary cells and MMC in 3 primary cells. Transplanted them to nude mice, experiment was divided into 3 parts(drug sensitivity test, traditional therapy and 0.9% natrii chloride). Chemotherapy filtered by drug sensitivity test in vitro were moreeffective than not filtered(p=0.034).ConclusionChemotherapy filtered by drug sensitivity test in vitro was more effective than not filtered. Part IVInhibition of PC Cell-derived Growth Factor Expression by shRNA transfection Increases Sensitivity of Pancreatic Cancer Cell linesPanc-1 to Gemcitabine ObjectPancreatic cancer is the most malignant disease of cancer. Most of patient lost chance of operation when they were diagnosed pancreatic cancer. More and more researchers are interesting about chemotherapy with gemcitabine. To detect the effect of PCDGF on sensitivity of pancreatic cancer cell lines Panc-1 to Gemcitabine by observing the changes after transfected. Methods and materialsWe investigated the expression of PCDGF in pancreatic cancer tissues and noncarcinoma tissues immunohistochemical staining and quantitative reverse transcription PCR(t=11.41, p<0.05); Two sites of shRNA targeted to PCDGF were combined to pGenesil-1 vector, and transfected to Panc-1 with lipofectamine2000, and measured the sensitivity of cancer cell lines to gemcitabine. ResultPCDGF was more expressed in pancreatic cancer tissues than pancreatic cytoadenoma and necrosis tissues in pancreatitis; Transfection shRNA targeted to PCDGF into Panc-1 cells could inhibit PCDGF expression (inhibit rate=82.3%). After transfection, the toxicity of gemcitabine to Panc-1 were remarkably increased especially in high concentration. ConclusionshRNA target to PCDGF make pancreatic cancer cell lines Panc-1 more sense to gemcitabine.