Ovarian cancer diagnosis or prognosis composition and application thereof
A technology of ovarian cancer and composition, applied in the field of biological diagnosis, to achieve the effect of increasing risk
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Embodiment 1
[0064] High expression of PDK1, PRKAA2, PRKAB1 mRNA in ovarian cancer cell line (SK-OV-3) and ovarian cancer tissues
[0065] In order to explore the biological role of PDK1 in ovarian cancer cells, we detected the mRNA expressions of PDK1, PRKAA2, and PRKAB1 in the ovarian cancer cell line SK-OV-3 by real-time fluorescent quantitative PCR, and used the HFL cell line as a control.
[0066] In order to explore whether the expressions of ovarian cancer cells in vivo and ovarian cancer cells in vitro are consistent with PDK1, PRKAA2, and PRKAB1, we collected 17 ovarian cancer tissues and 15 ovarian benign tumor tissues, and analyzed the two groups of tissues by real-time fluorescent quantitative PCR. Perform mRNA detection.
[0067] (1) Reverse transcription reaction:
[0068] 20μl system: mix RNA (adjusted to 100μg-500μg) with 4μl 5×PrimeScript RT Master Mix, and make up to a total volume of 20μl with RNase-free water;
[0069] The reverse transcription conditions are 37°C for...
Embodiment 2
[0081] 2.1 Immunohistochemistry
[0082](1) Bake the specimen slices in an oven at 80°C for 40 minutes.
[0083] (2) The specimen slices were dewaxed twice in xylene, each time for 13 minutes; hydrated with gradient alcohol (100%, 100%, 90%, 80%, 70%), each time for 5 minutes, soaked in pure water twice, each time 5 minutes each time;
[0084] (3) High-pressure antigen retrieval: Take an appropriate amount of immunohistochemical antigen retrieval buffer (powder type citric acid method) (1500ml) that has been diluted into a working solution in a pressure cooker, place the pressure cooker on an induction cooker, and heat to boiling with high power. Adjust the power to medium (500-1000W), place the slices on the high temperature staining rack, put them into the boiling repair solution, cover the pot, buckle the pressure valve, and continue heating. Time 3 minutes when the pressure valve starts to blow air. Then take the pressure cooker away from the induction cooker, let it co...
Embodiment 3
[0150] Example 3 The relationship between the expression of PDK1, PRKAA2 and PRKAB1 and the survival of patients with ovarian cancer
[0151] 3.1 The relationship between the expression of PDK1 and the survival of patients with ovarian cancer
[0152] Group the patients according to the following grouping requirements, and count the survival time of the patients, the same below
[0153] The grouping requirements are as follows:
[0154] PDK1 low expression group: cases with (-) and (+) PDK1 expression detected by immunohistochemical method
[0155] PDK1 high expression group: cases with (++) and (+++) PDK1 expression detected by immunohistochemical method
[0156] The result is as Figure 8 , P=0.195, there was no significant difference in the survival time of ovarian cancer patients between the PDK1 low expression group and the PDK1 high expression group. It can be seen that the expression of PDK1 has no significant correlation with the survival and prognosis of ovarian c...
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