noxa mRNA and its encoded protein expression in non-small cell lung cancer -related research
|School||Kunming Medical College|
|Course||Clinical Laboratory Science|
|Keywords||noxa Non-small cell lung cancer RT-PCR mRNA Protein Apoptosis Terminal deoxynucleotide Nucleotide transferase -mediated dUTP nick end labeling method in situ Western blot technique p53|
Objective To investigate the noxa mRNA expression and function associated with lung cancer; lung tissue p53 in the regulation of relations noxa. Methods Real-time quantitative RT-PC.R and immunoblotting (Western Blot, WB) in 44 patients with lung cancer tissues, adjacent tissues and distant lung cancer tissues and 10 cases of benign lesions noxa mRNA and its encoded protein expression; Application WB encoded protein expression of p53. Secondly, using TUNEL assay and 20 patients with lung cancer tissues, adjacent tissues and distant cancer cell apoptosis state. Results 1 real-time fluorescent quantitative RT-PCR, noxa mRNA Using ABI's TaqMan Universal PCR Master Mix, primers and probes specific pilot program. Reaction system TaqMan Universal PCR system 12.5μl, Assays-on-demand TM sup> Gene Expression Test genotyping reagents 1.25u1 (included forward primer and reverse primers and TaqMan probe), cDNA 2.51 μl, nuclease-free water, 8.75 u1, total system 25u1. The amplification of GAPDH primers and probes in addition, the other system components as above. Amplification conditions: 50 ℃ 2 min; 95 ℃ 10 min; 95 ℃ 15 sec, 60 ℃ 1 min, 40 cycles. (2) using real-time fluorescence quantitative RT-PCR, noxa mRNA expression of noxa mRNA in cancer tissues, adjacent group, far benign lesions and lung cancer group were expressed: cancer 0.52 (0.31 to 0.80), cancer tissue 1.49 (0.51 to 2.56), far cancer 1.00 (0.52 to 1.54). The rank sum test analysis, noxa mRNA expression in the same case of cancer tissues, adjacent tissues and far between cancer tissue was significantly (x 2 sup> (H) = 14.65, P lt; 0.05), pairwise comparisons, the results show: noxa mRNA expression in tumor and adjacent, cancer and distant cancer differences were statistically significant, while in the adjacent tissues and distant tumor tissues showed no significant difference between the , suggesting that cancer adjacent tissues and far between organizations noxa mRNA expression is insignificant, but were higher than noxa mRNA expression in tumor tissues. And squamous cell carcinoma, adenocarcinoma and adenosquamous carcinoma cancer tissues, adjacent tissues and distant cancer tissues noxa mRNA expression levels showed no significant difference (x 2 sup> (H) = 1.446, p gt; 0.05; F = 0.976, p gt; 0.05; F = 3.082, P gt; 0.05). 3.NOXA protein assay cancer tissues, adjacent tissues and distant cancer tissue NOXA relative to GAPDH scanning area ratios were 0.77 (0.40 to 1.59), 1.49 (0.79 to 2.69) and 1.40 (0.75 to 1.98). For lung cancer tissues, adjacent tissues, benign lesions and lung cancer tissues distant tissue NOXA protein expression rank test, the results showed: NOXA protein expression in the difference between the groups was statistically significant (x 2 sup> (H) = 23.211, P lt; 0.05); further to do pairwise comparisons between groups, the results showed: NOXA protein expression in tumor and adjacent, cancer and distant cancer, lung cancer and benign lesions were differences There was statistically significant (F = 9.064, P lt; 0.05), while in adjacent tissues, far cancer tissues and benign lesions were no significant differences between. Squamous cell carcinoma, adenocarcinoma and adenosquamous carcinoma cancer tissues, adjacent tissues and cancer tissues NOXA far no significant difference in expression levels (x 2 sup> (H) = 1.137, P gt; 0.05; x 2 sup> (H) = 0.000, P gt; 0.05; x 2 sup> (H) = 1.054, P gt; 0.05), shows that the expression level of NOXA histological type of lung cancer irrelevant. 4.P53 protein assay cancer tissue and cancer tissue, much cancerous tissue between benign lesions and lung cancer and adjacent tissues and benign lung lesions between p53 protein expression difference was statistically significant, while the rest was no difference between groups statistically significant. 5.TUNEL test results cancer tissues, adjacent tissues and distant cancer tissue A worker were: 4.96% ± 1.63% 1.45% ± 0.46,1.26% ± 0.42%. AI cancerous tissue is far higher than the adjacent tissues and cancer tissues, AI difference was statistically significant, Z = 3.920, p = 0.000; while adjacent tissues and far between cancer tissue AI also statistically significant difference, Z = - 3.469, P = 0.001. 6.p53 Relationship with NOXA NOXA protein expression and protein expression of P53 no direct relationship between, which may be lung cancer is due to a high of P53 mutation, mutant P53 lose NOXA role in regulating . Conclusion noxa mRNA, protein expression in lung cancer is lower, it may be associated with the occurrence and development of lung cancer is an important factor, but levels of mRNA and protein expression in lung cancer tissue types and pathological types; lung cancer NOXA and P53 no correlation between, suggesting that in this group of subjects is not mediated NOXA P53 expression may be the result of high mutation P53; cancer tissue increased apoptosis, NOXA expression is not the result of DNA damage may be other mechanisms cause. In addition, the findings in other tumor findings are not consistent, indicating noxa in tumor development, different organizations have different performance, and in tumor development mechanism of action remains to be further studied whether cancer diagnosis and treatment can be used as a new target remains to be further studied.