| Objective: The morbidity and mortality of lung cancer, which seriously threats human life and health, is the highest in malignant tumors. The non-small cell lung cancer(NSCLC) accounted for about 85% in lung cancer and five year survival rate of that was only 10%-15%. Owing to lack of obvious symptoms in early lung cancer, invasion and metastasis have been occurred before diagnosis and is an important factor for lung cancer patients survival. It is very important for the treatment of lung cancer that new drugs and effect targets which can inhibit lung cancer invasion and metastasis are found.The invasion and metastasis of lung cancer is a complex process and involves multi-factors and multi-signaling pathways influenced each other. To inhibit that, finding specific biomarkers and interfere expression is an effective way. Be a kind of calcium-dependent transmembrane glycoprotein, Epithelium cadherin(E-cadherin) mediates mutual epithelial cell adhesion, regulates the adhesion between cell and cell,as well as between cell and matrix, and plays an important role in proliferation, invasion and metastasis, and other links in non-small cell lung cancer. Ezrin is a membrane-cytoskeleton connection protein, which has no significant relationship with histological types, cell differentiation, clinical stage, but close relationship with lymph metastasis of lung cancer. Phosphatase and tensin homology deleted on chromosome ten(PTEN) is the first dual specific phosphatase activity tumor suppressor gene. Studies showed that the survival time of lung cancer patients with PTEN protein low or no expression were shorter than those with PTEN protein high or positive expression and occurrence of lymph metastasis were more higher. Cortactin is a microfilament actin-binding protein, which involves in tumor invasion and metastasis by influencing tumor cell motility, extracellular matrix degradation, anoikis, cell adhesion and other mechanisms. The high expression of Cortactin protein in lung cancer is closely related with its lymph node metastasis and clinical stage.Studies showed that cyclooxygenase-2(COX-2) was highly expressed in lung cancer and related with lung cancer cell proliferation, apoptosis, angiogenesis, lymph node metastasis and COX-2 inhibitors, such as celecoxib, nimesulide, inhibited lung cancer cell proliferation and angiogenesis and lymphangiogenesis, induced apoptosis, then to inhibit lung cancer invasion and metastasis. As a novel moderately selective COX-2 inhibitors, Imrecoxib can reduce serious cardiovascular events by highly selective COX-2 inhibitor. There is almost no reports about Imrecoxib can inhibit tumor invasion and metastasis at home and abroad. Lobaplatin, as the third generation platinum drugs, can block tumor cell DNA replication and transcription by the Pt-GG and Pt-AG cross connection within chains. Lobaplatin has superiority in significantly reducing the nephrotoxicity of cisplatin and the neurotoxicity of oxaliplatin. And it has broad-anti-tumor-spectrum, high concentrations in tumor tissue and low in plasma concentrations.Nowadays, it is unclear at home and abroad whether COX-2 inhibitors combined with chemotherapy drugs can enhance the efficacy of chemotherapy. The aim of study is to investigate whether Imrecoxib monotherapy or it combined with Lobaplatin can inhibit the growth of tumors in lung adenocarcinoma(A549) xenograft in nude mice and lung cancer invasion and metastasis, then to find mechanism with influence on E-cadherin, Ezrin, Cortactin and PTEN protein and m RNA corresponding expression.Methods:30 BALB / c nude mice, male, 4 ~ 5 weeks old, weighing 20 ~ 24 grams were selected and raised in SPF environment. Single cell suspension of logarithmic lung adenocarcinoma A549 cells cultured in F12 k medium was inoculated into the right axilla subcutaneously of nude mice to establish nude-mice transplanted tumor models of lung adenocarcinoma(A549) xenograft. The xenograft model meant success that the tumor average diameter reached 4mm. 1 nude mouse was removed which failed to forming xenograft model. 29 mice were randomly divided into four groups: control group, Imrecoxib group, Lobaplatin group, Imrecoxib combined with Lobaplatin group, Imrecoxib group had 8, other group had 7 nude mice. The control group was treated with the same amount of sterile distilled water and injected the same amount of saline by caudal vein, Imrecoxib group was treated with Imrecoxib tablets 40 mg.kg-1.d-1 through oral gavage while injected the same amount of saline by caudal vein, Lobaplatin group was injected Lobaplatin 7.5mg.kg-1.w-1 by caudal vein while the same amount of sterile distilled water through oral gavage, Imrecoxib combined with Lobaplatin group was treated with the same dose of Imrecoxib and Lobaplatin. After 6 weeks, the mice were killed by cervical dislocation and transplanted tumor tissues were cut. Caliper was used to measure the long diameter of tumor tissues(a) and the short diameter(b), according to the formula V=ab2/2 to estimate the approximate volume of tumor tissues and tumor tissues were weighted. Tumor tissues were fixed with 4% paraformaldehyde. E-cadherin, Ezrin, PTEN and Cortactin protein expression was detected with immunohistochemistry and flow cytometry. E-cadherin, Ezrin, PTEN and Cortactin m RNA content was detected with Real-Time PCR in each group tumor tissues.Results:1 29 nude mice were successfully formed transplant tumor in 30 nude mice. The time of tumor growth was 7-12 days after inoculating lung adenocarcinoma A549 cells. 1 nude mouse was died by oral gavage in Imrecoxib group. Compared with the control group, tumor volume is smaller(P<0.05), weight is less(P<0.05), necrotic area is more in Imrecoxib group, Lobaplatin group and Imrecoxib combined with Lobaplatin group; compared with the Imrecoxib or Lobaplatin group, tumor volume is smaller(P<0.05), weight is less(P<0.05), necrotic area is more in Imrecoxib combined with Lobaplatin group.2 Detected the expression levels of E-cadherin, Ezrin, PTEN and Cortactin protein in each group of lung adenocarcinoma(A549) xenograft in nude mice with immunohistochemistry and flow cytometry showed that, compared with the control group, the expression levels of E-cadherin and PTEN protein were significantly increased(P<0.05), the expression levels of Ezrin and Cortactin protein were significantly decreased(P<0.05) in Imrecoxib group and Imrecoxib combined with Lobaplatin group. Compared with the Imrecoxib or Lobaplatin group, the expression levels of E-cadherin and PTEN protein were significantly increased(P<0.05), the expression levels of Ezrin and Cortactin protein were significantly decreased(P<0.05) in Imrecoxib combined with Lobaplatin group. There was no statistically significant difference between Imrecoxib group and Lobaplatin group( P>0.05).3 Detected the contents of E-cadherin, Ezrin, PTEN and Cortactin m RNA with RT-PCR in each group of lung adenocarcinoma(A549) xenograft in nude mice showed that, compared with the control group, the contents of E-cadherin and PTEN m RNA were significantly increased(P<0.05), the contents of Ezrin and Cortactin m RNA were significantly decreased(P<0.05) in Imrecoxib group and Imrecoxib combined with Lobaplatin group. Compared with the Imrecoxib or Lobaplatin group, the contents of E-cadherin and PTEN m RNA were significantly increased(P<0.05), the contents of Ezrin and Cortactin m RNA were significantly decreased(P<0.05) in Imrecoxib combined with Lobaplatin group. There was no statistically significant difference between Imrecoxib group and Lobaplatin group( P>0.05).Conclusions:1 Imrecoxib or combined with Lobaplatin could inhibit the growth of tumors in non-small cell lung cancer.2 Imrecoxib or combined with Lobaplatin could upregulate the expression levels of E-cadherin protein and E-cadherin m RNA,PTEN protein and PTEN m RNA,could downregulate the expression levels of Ezrin and Ezrin m RNA, Cortactin protein and Cortactin m RNA to inhibit non-small cell lung cancer invasion and metastasis; Imrecoxib combined with Lobaplatin could enhance the effects on inhibition non-small cell lung cancer invasion and metastasis than single Imrecoxib or Lobaplatin, Imrecoxib could have synergistic effect on Lobaplatin chemotherapy.3 Imrecoxib may become a new target for inhibiting lung cancer invasion and metastasis, and a breakthrough in lung cancer treatment. |
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