Application of NY-ESO-1 in diagnosis and treatment of microsatellite instability intestinal cancer
A NY-ESO-1, instability technology, applied in the fields of biology and medicine, can solve problems such as not recommending chemotherapy and microsatellite instability
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Embodiment 1
[0113] Example 1 Clinical retrospective analysis of the impact of NY-ESO-1 expression differences on the survival time of patients under different microsatellite status
[0114] In this example, 186 clinical samples were collected from 2005 to 2009, and their microsatellite status was determined through microsatellite typing analysis. Among them, 123 cases of MSS and 63 cases of MSI-H patients were analyzed by immunohistochemical method. The expression of NY-ESO-1 in tissues showed that the proportion of positive expression of NY-ESO-1 in MSI-H was 57.14%, which was significantly higher than that in MSS.
[0115] The influence of NY-ESO-1 expression differences in different microsatellite states on the survival time of patients was analyzed retrospectively. In the MSI-H group, the three-year survival rate of patients with positive NY-ESO-1 expression was 74.18%, while the three-year survival rate of patients with negative NY-ESO-1 expression was 55.71% figure 1 A); while in t...
Embodiment 2
[0116] Example 2 Verification of high expression of NY-ESO-1 in MSI colorectal cancer
[0117] Adherent tumor cells were digested with trypsin, collected by centrifugation and washed twice with PBS, 1×10 6 Cells, RNA extraction kit, extract total RNA from cells, and store at -80°C after final concentration and purity determination. Take 2 μg of total cellular RNA and synthesize cDNA by reverse transcription according to the instructions of the reverse transcription kit, react at 42°C for 30 minutes, inactivate at 99°C for 5 minutes, then add 16 μl of double distilled water to dilute, which can be used as a template for qPCR reaction. Use MAGE-A4, NY-ESO-1, NY-ESO-2 and Actin quantitative PCR primers (F: 5'-CAGACCACCGCCAACTGCA-3' (SEQ ID NO.: 3); R: 5'-TGAGGGAGGCTGAGCCAAA -3'(SEQ ID NO.:4)), using Green Realtime PCR Master Mix for quantitative PCR amplification. Such as figure 2 As shown in A.
[0118] Adherent tumors were digested with trypsin to make a single cell susp...
Embodiment 3
[0119] Example 3 Using NY-ESO-1 expression to classify intestinal cancer microsatellites
[0120] Double-blind determination was carried out by using the intestinal cancer specimens with known microsatellite typing in Example 1, so as to classify the intestinal cancer specimens according to the positive expression of NY-ESO-1. The results showed that the sensitivity of judging microsatellite instability colorectal cancer based on the expression of NY-ESO-1 was 90%, and the specificity was 78%. Therefore, colorectal cancer can be classified according to microsatellite stability according to the expression of NY-ESO-1.
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