| Background Lung cancer is one of the most common malignant tumor in the world, the mechanism of which is not yet clear.The microRNA34a family is a highly conserved microRNA family,is a tumor suppressor gene,while c-myc is a member of myc gene family, also is the target gene of microRNA34a. Repot about research of the microRNA34a/c-myc pathway in NSCLC tissue was rare.This subject detected the expression of paper microRNA34a, c-myc gene and protein in NSCLC tissue,adjacent mucosa tissue and normal lung tissue using q RT-PCR and western-blot on the basis of previous study.The study explored the relationship between the two and the influence to the occurrence and development of NSCLC to reveal the molecular mechanism of the occurrence and development of NSCLC and to provide new approaches for the diagnosis and gene therapy of NSCLC patients.Objective The project mainly used the method of real-time fluorescent quantitative PCR(q RT-PCR) and western blot to detect the expression of microRNA34a and c-myc in normal tissue and NSCLC (non-small cell lung cancer) respectively. Explore the regulatory mechanism between microRNA34a and c-myc and the relationship with the NSCLC.Methods All the samples were derived from the pathology department of Xin Xiang central hospital and the people’s liberation army371hospital from April2012to June2013. Keep the NSCLC tissue, adjacent mucosa (3-5cm outside the cancer tissue) and normal lung tissue in-80℃freezer or liquid nitrogen. The NSCLC tissue, adjacent mucosa and normal lung tissue were all diagnosed or classified by pathological findings. The collected cancer patients excluded recurrent NSCLC patients, or NSCLC patients after chemotherapy before surgery.Use real-time fluorescent quantitative PCR method and Western blot method to detect the expression of microRNA34a and c-myc gene, c-myc protein respectively. Relevant analysis such as t-test and one-way analysis of variance were carried out using the statistical software Graph pad5.0.Result1. In the24cases of lung cancer tissue samples, there were16cases of lung squamous cell carcinoma,8cases of adenocarcinoma,12cases of adjacent mucosa and12cases of normal lung tissue.2. The expression results of microRNA34a by real-time fluorescent quantitative PCR are as follows：control grou0.918±0.096;adjacent mucosa group0.172±0.040; NSCLC group0.044±0.0310; There were significant differences between the experimental group and control group (P<0.01). There were significant differences in the experimental groups (P<0.01).3. The expression results of c-myc by real-time fluorescent quantitative PCR are as follows:control group0.364±0.086; adjacent mucosa group0.713±0.095; NSCLC group1.43±10.150; There are significant differences between the experimental group and control group (P<0.01), the differences between the experimental group, each group (P<0.01).4. The gray value analysis results of c-myc protein by western-blot are as follows: controlgroup34.35±4.869;adjacent mucosa group68.12±7.398;NSCLC group107.9±1.694; Compared with adjacent mucosa group, the gray value of the the NSCLC group increased statistically (P<0.01). Compared with the normal tissue group, the gray value of the NSCLC group increased statistically (P<0.01). There were significant differences in the experimental groups(P<0.01).5. Correlation between gene and protein expression of microRNA34a and c-myc, microRNA34a and c-myc gene both showed a negative correlation in all stage NSCLC tissue by real-time fluorescent quantitative PCR.the microRNA34a gene and c-myc protein by western-blot also showed a negative correlation, c-myc gene and c-myc protein were positively correlated, accorded with consistency analysis.Conclusion1. MicroRNA34a and c-myc showed a negative correlation in the development process of NSCLC, and had a very close inner relationship with the evolution of NSCLC.2. The abnormal control loop of microRNA34a/c-myc in NSCLC tissues provided new ways and ideas for early diagnosis and clinical therapy for NSCLC. |
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