| Background:Lung cancer is a common malignant tumor in death worldwide moorbidity and mortality is high,at present the pathogenesis of lung cancer is unclear.MicroRNA-21is a kind of small noncoding mas, participating in the regulation of gene transcription level after, in a variety of expression in tumor tissues such as esophageal cancer, gastric cancer, ovarian cancer and so on. Caspases-3is contain cysteine aspartic acid proteolytic enzymes can target specific cutting eggs Bai Tiandong glycine base sfter the peptide bond,caused by cell apoptosis,and microRNA-21of target genes.Abmormal pathways regulating its significance in the development of lung cancer,not reported at present.This topic by real-time fluorescent quantitative PCR (qRT-PCR) detection, such as detection of mirnas in rat lung squamous microRNA-21, the expression of Caspases-3explores the microRNA-21/Caspases-3pathway abnormality impact on development of rat lung squamous carcinoma and provides new train of thought and the scientific basis for clinical prevention and treatment of lung cancer.Objective:The detection the expression of microRNA21and Caspases-3in the development process of lung cancer in rat. Explore the role and significance between microRNA21and Caspases-3and lung cancer.Methods:1. Rats lung cancer model:the experimental group (60) with3-Methylcholanthrene (MCA) and Diethyl nitrosamine (DEN) lipiodol solution0.1ml in the left lower lobe bronchial perfusion; the control group (8) perfusion0.1ml lipiodol. To get lung squamous cell carcinoma of the formation of the specimens of the various stage of the process, HE staining, microscopic dianosis typing.2. Real-time PCR was used to detect microRNA21expression in the various stages of the diseased tissues.3. The reverse transcription-polymerase chain reaction (RT-PCR) technique was used to detect PTEN mRNA expression in lung squamous cell carcinoma and in control group.4. The expression of PTEN was examined by immunohistochemical technique.5. These results were statistically analyzed using SPSS16.0package.Results:1. Animal models48Wistar rats in the experimental group cancerous induced cancer was80%(48/60), specimens of cancerous lesions coexist with multiple stages, to obtain a total access to the bronchial epithelium hyperplasia tissue21cases,13cases of atypical hyperplasia, the original bit carcinoma in28cases,20cases of invasive cancer,16cases of metastatic cancer.8Wistar rats in the control group, no tumors occurred;2. MicroRNA21expression The expression results of microRNA21by real-time PCR are as follows:control group0.0017±0.0006; the atypical hyperplasia0.0178±0.0055; invasive group0.0589±0.0036; metastatic cancer group0.1327±0.1340. There are differences between the control group and the experimental group (.P<0.05), the differences between the experimental group, each group (P<0.05);3. Caspases-3protein and its gene expression According to the immunohistochemistry score Caspaces-3protein expression in different stages of ratings results were as follows:control group,4.50±0.93; atypical hyperplasia group,2.80±1.26; invasive group,2.00±1.43; metastatic cancer group,1.00±0.78. Compared the experimental group and the control group, the difference was statistically significant (P<0.01), the difference between atypical hyperplasia group and invasive group metastatic cancer group group also has a significant (P<0.01), the expression of Caspaces-3protein in the cancer tissues were lower than the atypical hyperplasia group and the control group (P<0.01). The analysis results of Caspases-3mRNA were as follows:control group,1.1217±0.0165; atypical hyperplasia group,1.0350±0.0320; invasive group,0.7000±0.0209; metastatic cancer group,0.4891±0.0133. Compared the experimental group and the control group, the difference was statistically significant (P<0.01), the difference between atypical hyperplasia group and invasive group, metastatic cancer group group also has a significant (P<0.01), the expression of Caspaces-3protein in the cancer tissues were lower than the atypical hyperplasia group and the control group (P<0.01).Protein and gene expression are consistent (Kappa=0.609, P<0.01);4. Correlation between microRNA21and Caspases-3In lung squamous cell carcinoma, microRNA21and Caspases-3protein both showed a negative correlation (r=-0.910, P=0.000); microRNA21and Caspases-3mRNA both showed a negative correlation (r=-0.953, P<0.01).Conclusions:1. The content of microRNA21in atypical hyperplasia, invasive carcinoma, and metastatic carcinoma tissues were higher than normal tissues; Caspaces-3were lower than normal tissues. MicroRNA21and Caspases-3showed a negative correlation In lung squamous cell carcinoma, they have a close relationship with the occurrence of lung cancer development.2. MicroRNA21and Caspases-3have certain internal relations, the joint detection of early diagnosis of lung cancer, as the treatment and prognosis monitoring indicators in lung cancer. |
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