Application of quantitative detection of CPT1A gene or protein in prognosis of esophageal squamous cell carcinomas
A quantitative detection, esophageal squamous cell carcinoma technology, applied in the direction of microbial determination/inspection, measuring devices, instruments, etc., can solve the problems of failure to apply clinical detection
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Embodiment 1
[0074] Retrospective study: DNA was extracted from 151 esophageal squamous cell carcinoma specimens (from Cancer Hospital, Chinese Academy of Medical Sciences), and the copy number of CPT1A gene was detected by Real-time PCR method and analyzed.
[0075] DNA copy number analysis: DNeasy Blood & Tissue Kit (Qiagen) kit was used to extract tumor tissue DNA according to the instructions, and the tumor cell content in the tumor tissue was required to be greater than 50%. Using Power SYBR The Green PCR Master Mix (Applied Biosystems) kit was used for real-time quantitative PCR (Real-time PCR) detection. The reaction system is 20μl: 10μl 2×Power SYBR Green PCR Master Mix, 2 μl of tumor tissue DNA (5ng / μl), 1 μl of primers (upper and downstream primers each 10 μM), 7 μl of nuclease-free water.
[0076] The reaction program is: 95°C for 10 minutes; 95°C for 15 seconds, 60°C for 1 minute, 40 cycles.
[0077] Real-time PCR results using 2 -ΔΔCt The internal reference gene was GAPD...
Embodiment 2
[0086] Retrospective study: DNA was extracted from 84 esophageal squamous cell carcinoma specimens (from Lin County, Henan Province), and the copy number of CPT1A gene was detected and analyzed by Real-time PCR method. The DNA copy number analysis method is the same as in Example 1. The result is as figure 2 and shown in Table 2.
[0087] Table 2. Multivariate regression analysis
[0088]
[0089] It can be seen from the results that patients with increased copy number (gain) of the CPT1A gene have a shorter postoperative survival time than patients without gain.
Embodiment 3
[0091] Retrospective study: 73 cases of esophageal squamous cell carcinoma tumor specimens were prepared for tissue microarray, and the protein expression level of CPT1A gene was detected and analyzed by immunohistochemical method.
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