Application of miR-133b in diagnosing, forecasting and treating prostate cancer with osseous metastasis
1. The technology of mir-133b and prostate cancer, which is applied in the field of molecular biology, can solve the problems affecting the treatment effect of tumors and the absence of miR-133b.
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experiment example 1
[0026] Experimental example 1. miR-133b is lowly expressed in prostate cancer tissue, and further lowly expressed in bone metastatic prostate cancer tissue
[0027] method:
[0028] RNA extraction, reverse transcription and real-time quantitative RT-PCR detection of miR-133b: extract the total RNA of all tissues and cells according to the instructions of RNA extraction reagent (Qiagen Company), and perform reverse transcription according to the instructions of Revert Aid First Strand cDNA SynthesisKit (Thermo Fisher Company) , using iQ SYBR Green reagent and CFX96 real-time quantitative PCR system (BIO-RAD company), using 2-ddCt relative quantitative method for fluorescence quantitative detection, using U6 as internal reference, to calculate the relative expression of miR-133b, miR-133b and The primers for U6 were designed, synthesized and purified by Guangzhou Ruibo Biotechnology Co., Ltd.
[0029] result:
[0030] The results of miR-133b expression in clinical specimens an...
experiment example 2
[0031] Experimental example 2. Low expression of miR-133b is associated with bone metastasis status and bone metastasis-free survival time in prostate cancer patients
[0032]According to the bone metastasis status of clinical specimens, the correlation analysis was carried out combined with the detected miR-133b expression level, and the patients were divided into miR-133b high expression group and miR-133b low expression group according to the median value of miR-133b expression level in prostate cancer tissues The analysis results are shown in Table 1, and the expression of miR-133b is correlated with the bone metastasis of prostate cancer.
[0033] Table 1. Correlation between miR-133b expression and bone metastasis status in prostate cancer patients
[0034]
[0035] According to the follow-up data of clinical specimens, the correlation between the expression of miR-133b and the 5-year overall survival time and 5-year bone metastasis-free survival time of prostate canc...
experiment example 3
[0036] Experimental example 3. Overexpression of miR-133b can inhibit the migration and invasion of prostate cancer in vitro
[0037] method:
[0038] The human miR-133b (miRBase number MIMAT0000770) gene was cloned into the pMSCV-puro retroviral vector (Clontech Company) to obtain the miR-133b expression plasmid, and the negative control plasmid or miR-133b expression plasmid were transfected into PC-3 In cells, C4-2B cells and VCap cells, Transwell chamber method was used to detect cell migration and invasion, and fluorescence quantitative detection was used to detect the expression of miR-133b.
[0039] result:
[0040] The constructed miR-133b expression plasmid was transfected into PC-3 cells, C4-2B cells and VCap cells, and the expression level of miR-133b image 3 As shown, the miR-133b expression plasmid can up-regulate the expression of miR-133b, which can be used for further study.
[0041] The results of cell migration and invasion experiments were as follows: ...
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