| Bladder cancer is a malignant tumor that occurs in the epithelium of the urinary bladder,it is a common tumor in urinary system. It ranks ninth in the world in the incidence of malignant tumors. The biological characteristics of the high recurrence and metastasis of the patients greatly shorten the survival time of the patients,a serious threat to human health. Looking for more effective chemotherapy drugs, Can not only improve the efficiency of patients who can not be operated,but also for the application of postoperative adjuvant therapy in reducing relapse in patients with metastasis and improve prognosis aspect provides a more effective solution. This experiment adopts mitomycin joint less research at home and abroad, beauty nimesulide as the research object, In bladder cancer EJ cells as experimental object, this paper discusses the combination of efficient.Mitomycin were adopted from the last century, the generation of chemotherapy drugs, antibiotic chemotherapy drugs, By Streptomyces extraction, chemical structure with benzoquinone, acetyl amino and carbamyl three active group, role similar to alkylating agent, With DNA cross-linking, inhibit DNA replication and RNA has inhibitory effect, thus inhibiting cell mitosis process, anti tumor formation. The cell cycle non-specific drug. Mainly used for a variety of solid tumors, such as stomach cancer, colon cancer, liver cancer, pancreatic cancer, non-small cell lung cancer, breast cancer and carcinoma of the breast, ascites, etc. Its treatment effect is clear, is the superficial bladder cancer surgery perfusion chemotherapy and late recurrence of bladder cancer metastasis and can’t surgery in patients with systemic chemotherapy is one of the commonly used chemotherapy drugs.Nimesulide, beauty is a kind of nonsteroidal anti-inflammatory drugs(nsaids), belongs to II ring oxidase inhibitors. The mechanism for Ⅱ can selectively inhibit an enzyme called cyclooxygenase, Related research in recent years show that II ring oxidase(cyclooxygenase 2, cox-2) in malignant tumor cells play an important role in the process of the growth, proliferation, it can promote tumor cell proliferation, inhibit tumor cell apoptosis, promote the tumor invasion and metastasis, and so on. For example, studies have found that central oxidase increased- 2 expression in bladder cancer cells, participate in the growth of tumor cell proliferation and apoptosis, and angiogenesis in tumor tissue, biological transfer play an important role in the process. Selective- 2 ring oxidase inhibitors such as celecoxib, beauty nimesulide can obviously inhibit the growth of bladder cancer cell proliferation, inducing apoptosis and inhibition of new blood vessels and lymphatic vessel formation, thus inhibiting the tumor invasion and metastasi, Another study confirmed that in ovarian cancer treatment in the study of in vivo and in vitro, selective- 2 ring oxidase inhibitors can also inhibit tumors-2 expression of the inner ring oxidase, reduce the expression of VEGF, reduce angiogenesis and promote cell apoptosis play a role of anti-tumor. For bladder cancer, therefore, the high expression of ring oxidase 2, can choose selective- 2 ring oxidase inhibitors to treat.Objective:This experiment mainly discusses his beauty nimesulide(optional)- 2 ring oxidase inhibitors combined mitomycin for bladder cancer EJ cell inhibitory effect. Application of the above two drugs on EJ cells in vitro, through the determination of caspase apoptosis related gene- 3 / BAX/BCL-2 on the EJ cells express, observe its effect on promoting apoptosis of bladder cancer EJ cells.Methods:1.Cell source: bladder cancer EJ cell lines(purchased from Shanghai beautiful porch biological technology co., LTD.), drug cultivation time 24 h, 48 h, 72 h, respectively2.Randomized controlled principle was adopted in the experiment group: control group and experimental group and control group in which don’t do deal with cell culture as a negative control, the experimental group according to the experimental drug divided into 3 groups: mitomycin C group, the concentration of 2 mg/L; Beauty nimesulide group, concentration of 300 umol/L; Mitomycin C(2 mg/L), beauty nimesulide group(300 umol/L).3.The control group and experimental group training for 24 h, 48 h, 72 h, Q- PCR method measuring the Bcl- 2 mrna, the intervention group cells Baxm RNA and Caspase3 m RNA expression difference4.The control group and experimental group training for 24 h, 48 h, 72 h, Western blot method of each intervention group cell the Bcl- 2 protein, Bax protein and Caspase3 protein expression differences5. Statistical analysis: use SPSS19.0 experimental data, with mean + /-standard deviation(+ /- s) data. Comparison between multiple sets of use I * J two-factor factorial design data of variance analysis, the comparison between the two groups using SNK method, P < 0.01, said the difference was statistically significant.Result1. Q- PCR method in the experimental group and blank control group in bladder cancer EJ cells measures the expression of Bcl- 2 mrna and Baxm RNA and Caspas3 m RNA:after treated 24 h,48h,72 h, Compared with the blank control group, The expression of Bcl-2m RNA in each experimental group were decreased.Baxm RNA and Caspase3 m RNA all improved. The difference was statistically significant. Compared with the single drug group, The expression of Bcl-2m RNA was the lowest in the combined group,and the expression of Baxm RNA and Caspase3 m RNA was the highest( P < 0.01) The difference wasstatistically significant. And with the extension of the time of action, The expression of Bcl-2m RNA was gradually decreased in the combined group,the expression of Baxm RNA and Caspase3 m RNA was gradually improved. When the 72 h function was the most obvious(P<0.01), The difference was statistically significant.2. Western-blot method in the experimental group and blank control group in bladder cancer EJ cells measures the expression of Bcl- 2 and Bax and Caspas3 protain:after treated 24 h,48h,72 h, Compared with the blank control group, The expression of Bcl-2 protain in each experimental group were decreased.Bax and Caspase3 protain all improved. The difference was statistically significant. Compared with the single drug group, The expression of Bcl-2 protain was the lowest in the combined group,and the expression of Bax and Caspase3 protain was the highest(P<0.05) The difference was statistically significant. And with the extension of the time of action, The expression of Bcl-2 protain was gradually decreased in the combined group,the expression of Bax and Caspase3 protain was gradually improved. When the 72 h function was the most obvious(P < 0.05), The difference was statistically significant.Conclusion:The combinition of mitomycin and nimesulide has synergistic effect in the growth inhibition and the promoting apoptosis of bladder cancer EJ cells.And The inhibitory effect is enhanced with the prolonging of time. The mechanism of action is related to the expression of apoptosis related genes Bcl-2, Bax and Caspase3. |
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