Non-small cell lung cancer (NSCLC) marker and its application
A technology of non-small cell lung cancer and markers, applied in the direction of drug combination, genetic material composition, biochemical equipment and methods, etc., can solve the problems of no literature reports
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Embodiment 1
[0105] Example 1: Expression of STAT3 protein in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5
[0106] The results of immunohistochemical staining showed that the positive expression was brownish-yellow or brown granular and deposited on the positive site. The positive expression signal of STAT3 protein in human lung adenocarcinoma cell A549 is mainly located in the cytoplasm, and a few are located in the nucleus, showing obvious brownish yellow or brown granular staining (see attached figure 1 ). In the normal human embryonic lung cell MRC-5, it is also weakly expressed in the cytoplasm, showing light yellow coloration (see attached figure 2 ). The t-test was performed between the MOD values expressed in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5, P=0.006 (P<0.01), and the difference was statistically significant, suggesting that STAT3 protein was in The expression level in human lung adenocarcinoma c...
Embodiment 2
[0110] Example 2: Correlation between the expression of STAT3 protein and CEA protein and CA125 protein in human lung adenocarcinoma cell A549
[0111] 2.1 Expression of CEA protein in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5
[0112] The positive expression signal of CEA protein in human lung adenocarcinoma cell A549 is mainly located in the cytoplasm and (or) the cell membrane, showing brownish-yellow or yellowish-brown granular staining (see attached image 3 ); normal human embryonic lung cell MRC-5 was also expressed to a weaker degree in the cytoplasm, showing light yellow coloration (see attached Figure 4 ). The MOD value of this protein expressed in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5 was tested by t test, P=0.000 (P Figure 7 ,table 3).
[0113] Table 3 CEA protein expression in A549 and MRC-5 cells ( ±s)
[0114]
[0115] Note: a P<0.01 vs MRC-5
[0116] 2.2 Expression of CA12...
Embodiment 3
[0127] Example 3: Expression of STAT3 mRNA in human lung adenocarcinoma cell A549 and normal human embryonic lung cell MRC-5 In this experiment, the SYBR Green I fluorescent dye method was selected, and the melting curve analysis step was added to the experimental design, which was analyzed by Bio-Rad IQ5 Software analysis. Melting curves showed that each gene had a single product peak (such as Figure 8-1 , 8-2, 8-3 and 8-4), no non-specific products and primer-dimers were generated, indicating that the specificity of the primers was good, and the experimental results could accurately reflect the initial concentration of the sample.
[0128] To establish a standard curve: take a certain amount of cDNA PCR product template, and carry out 10 3 ~10 9 Dilute to obtain 7 concentration gradients, and then take 2 μl of dH 2 O as zero tube. At the same time, real-time PCR amplification was performed to establish a standard curve. The Ct value of the standard PCR product dilute...
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