Oral cavity squamous carcinoma diagnosis reagent, reagent kit and preventing and controlling medicament
A diagnostic reagent and technology for oral squamous cell carcinoma, which is applied in the field of tumor molecular biology, can solve the problems of drugs that have no oral squamous cell carcinoma, diagnostic reagents and kits that have no oral squamous cell carcinoma, and can improve the survival rate. , the effect of broad application prospects
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Embodiment 1
[0046] Example 1 Preliminary determination of proteins related to the occurrence and development of oral squamous cell carcinoma
[0047] In this experiment, the classic method of differential proteomics research was used to analyze the protein expression profiles of 20 cases of surgically resected fresh oral squamous cell carcinoma tissues and their adjacent normal tissues using two-dimensional electrophoresis-tandem mass spectrometry-bioinformatics as the core proteomics technology , high-throughput identification of differentially expressed proteins associated with tumors. The difference points were selected by the PDQuest analysis software and quantitatively analyzed, and the results were obtained: the protein spots separated by two-dimensional electrophoresis were analyzed by the difference software, and 134 difference points with repeated expression and a difference of more than 1.5 times were obtained. ESI-Q-TOF mass spectrometry successfully identified 120 protein spot...
Embodiment 2
[0048] Example 2 Further determination of RACK1 as a protein related to the occurrence and development of oral squamous cell carcinoma
[0049] 1. Cell line
[0050] Oral tongue cancer cell line Tca8113 was preserved by West China Stomatological Hospital, human immortalized oral keratinized epithelial cell line HOK16E6E7 was donated by Xuan Lin (Charles R.Drew Uni.of Medicine, USA)
[0051] 2. Main reagents
[0052] RT-PCR: AccessQuick TM RT-PCR System, purchased from Promega Company.
[0053] The RACK1 rabbit polyclonal antibody used in Western blotting and the goat anti-rabbit secondary antibody were purchased from Santa Crus.
[0054] 3. Extraction of total cellular RNA for one-step semi-quantitative RT-PCR
[0055] Equal amounts of total RNA from Tca8113 cells and HOK16E6E7 cells were extracted, and then one-step semi-quantitative RT-PCR was performed. The primer design is shown in Table 1, and the b-actin gene was used as an internal reference.
[0056] Table 1 One-...
Embodiment 3
[0063] Example 3 Differential expression of RACK1 in normal tissue, OLK tissue, and OSCC tissue
[0064] 1. Tissue samples and reagents
[0065] 48 cases of OLK (oral leukoplakia tissue) and 76 cases of OSCC paraffin specimens were collected from the pathology department of West China Stomatological Hospital in 2005-2006. The clinicopathological data were obtained from medical records and histopathological reports, as shown in Table 2-1. 20 cases were normal Tissues were discarded from oral and maxillofacial plastic surgery.
[0066] The RACK1 rabbit polyclonal antibody used in immunohistochemistry, the goat anti-rabbit secondary antibody was purchased from Santa Crus, and the SABC kit was purchased from Wuhan Boster Company.
[0067] 2. Immunohistochemical staining
[0068] All specimens were immunohistochemically stained by SABC method under the same conditions, and then SPSS 11.5 software was used to perform one-way analysis of variance for comparison between groups, and ...
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