Application of a lncRNA in the diagnosis and treatment of oral squamous cell carcinoma
An oral squamous cell carcinoma, a technology in the sample, applied in the field of biomedicine, can solve the problems of recurrence, metastasis, and the 5-year survival rate of patients has not been significantly improved.
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Embodiment 1
[0064] Example 1 Biomarkers associated with oral squamous cell carcinoma
[0065] A data set containing 372 samples (including 339 OSCC patient tissue samples and 33 normal tissue samples) was downloaded from the TCGA database, and the differential expression of IGF2BP2-AS1 was analyzed using the edgeR software package. Based on the expression of IGF2BP2AS1, the patients were divided into two groups with the optimal cut-off value, and the Kaplan-Meier method was used to construct the survival curve. A p value <0.05 was considered statistically significant.
[0066] Result: if figure 1 As shown in (a), the expression level of IGF2BP2-AS1 was significantly increased in OSCC patient tissues compared with normal tissues. In addition, if figure 1As shown in (b), the expression level of IGC2BP2-AS1 was negatively correlated with the survival of OSCC patients (P=0.031). The results of this experiment suggest that IGF2BP2-AS1 can be used for the diagnosis of OSCC and is associated...
Embodiment 2
[0067] Example 2 Expression of IGF2BP2-AS1 in OSCC tissues and cells.
[0068] 1. Sample acquisition
[0069] 30 OSCC tissues and 20 paracancerous tissues (more than 5cm away from the primary tumor) were collected from Shandong Provincial Hospital. The excised samples were all frozen at -80°C and confirmed by pathological diagnosis. The acquisition of all the above samples was approved by the Ethics Committee of Shandong Provincial Hospital.
[0070] 2. mRNA extraction and RT-qPCR
[0071] Firstly, the total RNA of the sample was extracted with TRIzol reagent, and then the RNA was reverse-transcribed to obtain cDNA using the HiFi-Script cDNA first-strand synthesis kit, and finally the UltraSYBR Mixture premix system was used for qPCR amplification, and the specific primer sequences were as follows:
[0072] IGF2BP2-AS1:
[0073] The forward primer is 5'-ATGGTGGTGCATGGAGGAAG-3' (SEQ ID NO.1);
[0074] The reverse primer is 5'-CTCTTCAGCAAAGCAGACTTC-3' (SEQ ID NO.2).
[0075...
Embodiment 3
[0084] Example 3 Silencing test and functional verification of IGF2BP2-AS1
[0085] 1. Cell culture
[0086] at 5% CO 2 1. Cells (Cal27, Tca8113) were cultured in a constant temperature incubator at 37° C., and the medium was DMEM medium containing 10% FBS.
[0087] 2. Cell transfection
[0088] The siRNA against IGF2BP2-AS1 was synthesized by Beijing Oligobio Co., Ltd. (Beijing, China), and its sequence is as follows:
[0089] si-IGF2BP2-AS1 is GGTCATGCCTGGTTAACAT (SEQ ID NO. 5).
[0090] The sequence of the siRNA used as a control is as follows:
[0091] si-NC is TTCTCCGAACGTGTCACGT (SEQ ID NO. 6).
[0092] The Lipofectamine 2000 kit was used for transfection, and the specific operation was carried out according to the instructions.
[0093] 3. Cell colony formation experiment
[0094] Cells (Cal27, Tca8113) were seeded into 35mm diameter culture dishes (1000 cells / well) and cultured for 14 days. Cells were fixed with 4% paraformaldehyde solution for 15 min and stain...
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