Research On The Effects And Reasons Of Nimesulide And Oxaliplatin On Human Lung Cancer Xenograft In Nude Mice

Objective: At present, due to a variety of factors such as smoking andenvironmental impact, the incidence of lung carcinoma is still very high.Although there are several treatments, the overall5-year disease free survivalrate is no noticeable difference. Lung cancer is the leading cause of death inthe word. Each year, about one million people, including850,000men and330,000women die of lung cancer. According to the histological classification,lung cancer includes two major subtypes, small cell lung cancer and non-smallcell lung cancer, the latter account for85%of all lung cancer cases, more than60%of patients with non-small cell lung cancer at the time of diagnosis is lateor metastatic, and no chance of surgery. Recently, the molecular targetedtherapy on lung cancer has became a research hotspot. Multiple genesinvolved in the invasion and metastasis of lung cancer. Survivin is animportant member of the inhibitor of apoptosis protein family, ananti-apoptotic protein, which has overexpressed in many tumors andprecancerous lesions. Inhibition of survivin expression can inhibit tumor cellproliferation and induce apoptosis. Matrix metalloproteinase-2, a member ofproteolytic enzymes, can degrade collagen type Ⅳand basement membranecomponents, which play a key role in the development of tumor invasion andmetastasis. CD44v6is a member of adhesion molecule family, and mediatecell-cell and cell-extracellular matrix adhesion effect.CD44v6belongs to themembrane surface glycoprotein, which generally express in human malignanttumors. Ki67is a cell proliferation related nuclear antigen. Its positiveexpression show cancer cells proliferation active. Nimesulide, selectivecyclooxygenase-2inhibitor, is an commonly used non-steroidalanti-inflammatory drug. The study confirmed that nimesulide can inhibittumor growth. Oxaliplatin is the third generarion drug of platinum chemotherapy medicine. To establish a human lung cancer xenograft on nudemice. This study is designed to evaluate the effects of non-steroidalanti-inflammatory drug nimesulide in combination with commonly usedoxaliplatin, on tumor growth, survivin, MMP-2, CD44v6and ki67expressionsin lung cancer xenograft on nude mice. To furtherly discuss the molecularmechanism. Committed to explore new effective drugs for the treatment oflung caner.Methods: Chose twenty-five four to five-week-old male nude mice. Bredunder specific conditions. Inoculated the human lung cancer A549cells in thelogarithmic phase in nude mice left axillary subcutaneous to establish humanlung cancer xenograft on nude mice.After the tumor diameter reached anaverage of4mm, removed the nude mouse which had the smallest tumorvolume and then randomly divided them into nimesulide-treated group,oxaliplatin-treated group, nimesulide combined with oxaliplatin-treated groupand control group. The oxaliplatin-treated group nude mice were injectedoxaliplatin mannitol injection,10mg/kg,drug delivery every three days.And atthe same time take sterile distilled water. The nimesulide-treated group wasadministrated nimesulide20mg/kg/d by intragastric administration.And at thesame time injected with equivalence physiological saline. The nimesulidecombined with oxaliplatin-treated group were administrated nimesulide20mg/kg/d by intragastric administration.And at the same time injected withequivalence oxaliplatin mannitol injection. The control group was treated withsterile distilled water, and at the same time injected intraperitoneally withphysiological saline. Administered thirty days later,the animals were sacrificed.Cut tumor tissue. The survivin, MMP-2, CD44v6and ki67protein expressionin the tumor tissue were detected by immumohistochemical method.Caculated the average integral optical density value by Buaa true colorpathological image analysis system. The survivin, MMP-2, CD44v6andki67mRNA expression in the tumor tissue were detected by florescentreal-time quantitative PCR.Results: Measured the integral optical density of survivin, MMP-2, CD44v6and ki67protein immunohistochemical staining sclices bypathological image analysis system. Compared with the control group,statistical analysis showed that the expression levels of survivin, MMP-2,CD44v6and ki67protein of nimesulide-treated group and nimesulidecombined with oxaliplatin-treated group were significantly reduced(respectively, P<0.05). Compared with the control group, the expression levelsof survivin and ki67protein of the oxaliplatin-treated group were significantlyreduced (respectively, P<0.05). Compared with the control group, theexpression levels of MMP-2and CD44v6protein of the oxaliplatin-treatedgroup have no difference (respectively, P>0.05). Compared with the othergroups, the survivin, MMP-2, CD44v6and ki67protein expression levels ofthe nimesulide combined with oxaliplatin-treated group were significantlyreduced (respectively, P <0.05).The survivin, MMP-2, CD44v6and ki67mRNA expression were detectedby fluorescent real-time PCR method. Compared with the control group,statistical analysis showed that the expression levels of survivin, MMP-2,CD44v6and ki67mRNA of nimesulide-treated group and nimesulidecombined with oxaliplatin-treated group were significantly reduced(respectively, P<0.05). Compared with the control group, the expression levelsof survivin and ki67mRNA of the oxaliplatin-treated group were significantlyreduced (respectively, P<0.05). Compared with the control group, theexpression levels of MMP-2and CD44v6mRNA of the oxaliplatin-treatedgroup have no difference (respectively, P>0.05). Compared with the othergroups, the survivin, MMP-2, CD44v6and ki67mRNA expression levels ofthe nimesulide combined with oxaliplatin-treated group were significantlyreduced (respectively, P <0.05).Conclusion: Using nimesulide or nimesulide combined with oxaliplatincould inhibit survivin, MMP-2, CD44v6and ki67expression. Usingoxaliplation can inhibit the expression of survivin and ki67. Combination drugof nimesulide and oxaliplation can enhance the anti-tumor efficacy.