Early diagnosis kit for liver cancer, and application of early diagnosis kit
An early diagnosis and kit technology, applied in the fields of biomedicine and molecular biology, to achieve the effects of good specificity, fast and convenient detection, and high detection sensitivity
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Embodiment 1
[0048] Example 1 Detection of the expression levels of HBXIP, miRNA-4784, and LncRNA RP11-624L4.1 in liver cancer and the establishment of a detection kit
[0049] 1. Collect 30 liver cancer patient tissues and adjacent tissues (5 cm from the tumor edge).
[0050] 2. RNA sample preparation: Extract tissue total RNA using TRIZOL method
[0051] 1) Lyse the tissue with 1ml Trizol reagent for every 50-100 mg of tissue; 2) Transfer the Trizol lysate of the above-mentioned tissue into an EP tube, and place it at 15-30°C for 5 minutes; 3) In the above-mentioned EP tube, Add 0.2ml of chloroform after every 1ml of Trizol, cover the EP tube cover, shake vigorously in your hand for 15 seconds, and place it at room temperature (15-30°C) for 2-3 minutes, then 12000g (2-8°C) Centrifuge for 15 minutes; 4) Remove the upper aqueous phase and put it in a new EP tube, add isopropanol according to the amount of 0.5ml isopropanol per 1ml Trizol, and place it at room temperature (15-30°C) for 10 ...
Embodiment 2
[0075] The detection of the clinical sample by the kit of embodiment 2
[0076] Using the detection kit of Example 2, the expression levels of HBXIP, miRNA-4784, LncRNA RP11-624L4.1 in the serum of 30 cases of liver cancer patients and 30 cases of healthy people (2 -△△CT value) to check. The detection results were analyzed using origin software. The results of single detection indicators, combination of two indicators and joint detection results of three indicators of HBXIP, miRNA-4784, LncRNA RP11-624L4.1 are shown in the table below.
[0077]
[0078] As shown in the table above, the P values of each index and the combined index were all <0.05, indicating that each index was significantly correlated with the prediction of liver cancer. The joint prediction accuracy of HBXIP / miRNA-4784 / LncRNA RP11-624L4.1 reached 98% (the area under the ROC curve), which was higher than the prediction rate of a single detection index / two joint detection data (both less than 90%). Accor...
Embodiment 3
[0079] Example 3: Preparation and verification of pharmaceutical composition for treating liver cancer
[0080] From the above two examples, it can be concluded that the combined detection of the expression levels of HBXIP, miRNA-4784, LncRNA RP11-624L4.1 can be used as a diagnostic marker for liver cancer, and in liver cancer tissues, HBXIP, miRNA-4784, LncRNA RP11-624L4 The expression level of .1 was significantly upregulated. We speculate that the use of HBXIP, miRNA-4784, and LncRNA RP11-624L4.1 interferers may have certain therapeutic effects on liver cancer, thus providing a new possibility and direction for targeted therapy of liver cancer. In view of this, we designed and synthesized si-HBXIP (see SEQ ID NO.1 for the specific sequence), si-miRNA-4784 (see SEQ ID NO.2 for the specific sequence), si-LncRNA RP11-624L4.1 (The specific sequence is shown in SEQID NO.3).
[0081] Nude mouse tumor model was used to verify the experimental treatment of related drugs, and the ...
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