Kit for colorectal cancer auxiliary diagnosis and/or prognosis
A technology for auxiliary diagnosis and prognosis judgment, applied in biological testing, biochemical equipment and methods, microbial measurement/testing, etc., can solve problems such as blocking tumor formation
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Embodiment 1
[0055] Embodiment 1, clinical sample collection
[0056] For the determination of mRNA content: a total of 98 colorectal cancer samples from February 2010 to August 2012 were selected from Sun Yat-Sen University Cancer Center, and fresh tissues were taken after surgery and stored in liquid nitrogen; for protein content determination samples: select A total of 210 colorectal cancer specimens were collected from Sun Yat-sen University Cancer Center from September 2000 to January 2006. The paraffin specimens were stored at room temperature, and the cut white slices were stored in a 4°C refrigerator.
[0057] For each selected case, the inclusion criteria were newly diagnosed, pathologically diagnosed as single colorectal adenocarcinoma, no preoperative radiotherapy and chemotherapy, and radical resection was required for TNM stage I, II, and III cases. Each patient contained tumors and normal controls (normal mucosa more than 5 cm from the resection margin).
[0058] For all cas...
Embodiment 2
[0059] Example 2, quantitative PCR detection of PCDHB3 mRNA levels in colorectal cancer tissues and cell lines
[0060] 1) Total RNA was extracted from 98 colorectal cancer tissues and their normal controls, 8 colorectal cancer cell lines and two normal control cell lines using the TRIZOL method, and 2 μg of total RNA was reverse-transcribed into cDNA.
[0061] 2) Query the Gene database of NCBI to obtain the cDNA sequences of PCDHB3 and GAPDH (internal reference), and design PCR primers according to the sequences as follows:
[0062]
[0063] 3) Establish the following reaction system, and set up three reaction wells for each sample:
[0064]
[0065] Mix the above components, add to each well of the 8-strip tube, close the lid tightly, remove air bubbles, and centrifuge to make the liquid gather to the bottom of the tube. The thermal cycle parameters were as follows: 95°C for 10 min, 95°C for 30 s, 60°C for 30 s, 72°C for 40 s, 40 cycles, 95°C for 1 min, 60°C for 30 s...
Embodiment 3
[0070] Example 3, Determination of PCDHB3 protein content in colorectal cancer tissue specimens
[0071] 1) During the experiment, 210 cases of colorectal cancer cases were taken out paraffin section white slices, dewaxed with xylene, hydrated with gradient alcohol, and 0.3% H 2 o 2 Detissue peroxidase, citrate solution microwave repair, Abgent anti-PCDHB3 (AP12159a) antibody (1:100) and corresponding species secondary antibody incubation, DAB and hematoxylin color development, hydrochloric acid alcohol differentiation, gradient alcohol dehydration After the xylene transparency step, the slides were sealed with neutral gum.
[0072] 2) The staining evaluation of tissue samples was taken by a 200× inverted microscope, and 5 visual fields / cases were taken for each of the paracancerous and cancerous tissues, and then the pictures were analyzed with inForm1.4.0 software. The staining intensity of PCDHB3 in cells was divided into four grades: negative (0), weak positive (1), mode...
PUM
![No PUM](https://static-eureka.patsnap.com/ssr/23.2.0/_nuxt/noPUMSmall.5c5f49c7.png)
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