Application of TMC5 in diagnosis and treatment of breast cancer specific bone metastasis
A specific, breast cancer technology, applied in the field of biomedicine, can solve the problems of invasive examination, limited sensitivity and specificity, and achieve the effect of inhibiting bone metastasis and inhibiting migration
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Embodiment 1
[0031] Example 1 Study on the expression of TMC5 in breast cancer tissue.
[0032] 1. Experimental materials: breast cancer tissue samples, adjacent breast cancer tissue samples, TRIzol, dd water, isopropanol, chloroform, etc. The detailed reagents are listed in the specific experimental methods.
[0033] 2. Experimental method.
[0034] 2.1 Tissue: Wipe the ophthalmic scissors and tweezers with alcohol, and thaw the cryopreservation tube containing the tissue in advance. After the ice cubes disappear, use the ophthalmic scissors to cut off 50-100 mg of tissue samples, rinse with dd water, and use de-enzyme water. The soaked filter paper squeezes out the oil or blood components in the tissue sample, and then immerses the sample into a 1.5mL EP tube containing TRIzol, and shreds it with ophthalmic scissors to make the TRIzol fully contact with the tissue sample to facilitate RNA extraction.
[0035] 2.2 Extracting total RNA, the specific steps are as follows:
(1) Let the mix...
Embodiment 2
[0045] Example 2 Study on the expression of TMC5 in breast cancer cell lines.
[0046] 1. Experimental materials: breast cancer cell lines (such as figure 2 Note), human normal breast cells, TRIzol, dd water, isopropanol, chloroform, etc.
[0047] 2. Experimental method.
[0048] 2.1 Cells: When the cells grow to 90% density, aspirate and discard the medium, wash twice, trypsinize for different times according to different cell lines, aspirate and discard the trypsin, pipet cells in complete medium, centrifuge in a 1.5mL EP tube, Discard the supernatant, add 1 mL of TRIzol, and blow well to make the TRIzol fully contact with the cells.
[0049] 2.2 Whole protein extraction from breast cancer tissue:
(1) Add 0.4 μL of phosphatase inhibitor, 0.2 μL of PPMSF and 0.04 μL of protease inhibitor to 40 μL of LysisBuffer to prepare a protein lysis solution;
(2) Aspirate the cell culture medium and wash it twice with PBS;
(3) Add 40 μL of lysis solution to each well of cells, p...
Embodiment 3
[0052] Example 3 Analysis of clinical case data The correlation between TMC5 and breast cancer-specific bone metastasis.
[0053] 1. Experimental materials: Collect patient data and follow up.
[0054] 2. Experimental methods: Chi-square test, Wilcoxon rank sum test, t test, Kaplan-Meier survival curve, etc. used in this part are based on SPSS statistical software (version 19.0, IBM, Chicago, IL, USA). For all test results, p < 0.05 was considered statistically significant.
[0055] 3. Experimental results: Through the analysis of the clinical case data of the patients, it was found that TMC5 was significantly correlated with breast cancer-specific bone metastases (Table 1). TMC5 was significantly and specifically highly expressed in breast cancer patients with bone metastases, but it was not found in liver and lung metastases.
[0056] Table 1: Clinicopathological data of 159 breast cancer patients included in the study
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[0057] * in the table means: p<0.05. a: Fish...
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