| Backgruond and objectiveBladder cancer is one the most common malignant tumor in China of all kinds of tumors, a major killer of human health, aseriously threaten to people’s health and lives and has has a high morbidity and mortality. Bladder cancer treatment mainly surgery (removal of the tumor tissue) in combination with other treatment (including intravesical chemotherapy, radiotherapy, chemotherapy, neoadjuvant chemotherapy), but even if there is a variety of treatment methods, the recurrence rate is still high and the tumor grading may be progression. So seriously affected the 5-year survival of patients and combat the confidence of related postoperative treatment. The main reason of relapse is surgery can not completely removed all of the tumor cells therefore an urgent need to explore new treatment options, both to reduce the risk of recurrence of bladder cancer, but also reduce resistance and reduce the side effects of drugs. In the treatment of bladder cancer, the immune therapy of tumors by stimulating or in combination the body’s immune system to strengthen the body’s immunity against tumors while mediated immune system to kill tumor cells removed. Raman spectroscopy as a simple, fast, non-destructive, highly sensitive detection method can analysis the molecular structure of the material by spectroscopic detection. Raman spectra of tissue or cell analysis changes in their internal bond vibrations associated components such as carbohydrates, proteins, lipids, nucleic acids and there internal components, structural changes it is possible to form a feature of the Raman peaks. We use bladder cancer-specific antibody (BCMabl) combined with cisplatin in vitro cell research, according to Raman spectrum to detecte molecular characteristics of conformational changes analysis of after different treatment methods the cells characteristics Raman peaks changes inferred bladder cancer-specific antibody(BCMab1) and cisplatin synergistic anti-cancer effect of bladder, explore the mechanism when bladder cancer-specific antibody (BCMabl) and cisplatin action to bladder cancer cell case to molecule changes then provide the basis for the study of anticancer drugs combination therapy.Materials and Methods:EJ bladder carcinoma cell line, hybridoma used for production antibodies (BCMabl) against human bladder cancer, Balb/c mice used for production tumor Ascites, and Cisplatin all provided by the Institute of Biophysics, Chinese Academy of Sciences(A) Firstly, use ascites made monoclonal antibodies, preparation preliminary antibodies, purified antibody, using flow cytometry to testing antibody 。(B) According to the design take on bladder cancer cell line EJ cells using coverslips plus six-well plates to culturing, the design:the control group (without drug treatment) Anti-alone group (BCMab1 final concentration of 25μg/ml and 50μg /ml) alone cisplatin (DDP final concentration of 25μg/ml and 50μg/ml) antibody mixed with cisplatin group (final concentration BCMabl,25μg/ml+DDP,25μg/ml; BCMabl,25μg/ml+DDP,50μg/ml; BCMabl,50μg/ml+DDP,25μg/ml; BCMabl,50μg/ml+DDP,25μg/ml) were cultured for 48h (3 replicates per group), using paraformaldehyde fixing Cells and imparity concentration ethanol to removing fixative and the water intracellular after the cells were fixed, Using 785nm excitation Raman spectroscopy gathering the Raman spectrum and references Raman bands attribution which have being summarized by the document, according to analysis the characterized by peaks significance, we can explore the molecular and biochemical changes in bladder cancer cell lines, The original resulting data are deal with the origin 8.6 Premier Edition。Results1. without any drug group, plus antibody alone group, plus cisplatin alone, cisplatin plus antibody mixing the drug group, cultured 48h average Raman spectra were 1450 cm-1,1700-1,1031 cm-1,1260 cm-1,1094 cm-1,1209 cm-1 showing a characteristic Raman peaks2. In accordance with the overall plan to join the pharmaceutical Raman spectral curves were higher than the negative control group3. Add antibody alone were higher than the overall Raman curve curves plus cisplatin alone4 antibody alone group to join, join cisplatin alone, cisplatin antibody mixed group, and no significant new features of the Raman peaksConclusion:1. As a non-specific cell cycle effects of cisplatin on apoptosis can be used Raman spectroscopy overall evaluation, but specific targets and detailed effects can not tell. And it can not produce its own action appears its unique specificity of Raman shift peak after Acting on bladder cancer EJ cells.2.antibodies against human bladder cancer (BCMabl) on apoptosis effect can be evaluated by Raman spectroscopy overall, but the specific targets, and detailed effects can not display. And it can not produce its own action appears its unique specificity of Raman shift peak after bladder cancer EJ cells.3. The antibody against human bladder cancer (BCMabl) the effect of cisplatin (DDP) combined effects after Acting on EJ cells can Raman spectroscopy overall evaluation, but specific targets and detailed effects can not display. And the combined effects of bladder cancer does not appear to produce its unique specificity of Raman shift peak after EJ cells.4.Raman bladder tissue by reference to past research found that the conclusions of bladder cancer EJ cell line also has a portion of the tissue specificity of Raman shift peaks.5. Through this experiment can prove that antibodies against human bladder cancer (BCMabl) and cisplatin (DDP) combined effects of EJ cells do not have obvious synergistic effect. |
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