Use of a piwi protein-interacting RNA piR-hsa-211106

A protein interaction and application technology, applied in the field of biomedicine, can solve problems such as the growth process of unseen lung adenocarcinoma

Active Publication Date: 2021-09-14
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no literatures or reports about piRNA regulating the growth process of lung adenocarcinoma in vivo or using piRNA that regulates the growth of lung adenocarcinoma to treat lung adenocarcinoma

Method used

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  • Use of a piwi protein-interacting RNA piR-hsa-211106
  • Use of a piwi protein-interacting RNA piR-hsa-211106
  • Use of a piwi protein-interacting RNA piR-hsa-211106

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example involves the process of constructing an agonist for PIWI protein-interacting RNA piR-hsa-211106: the PIWI protein-interacting RNA piR-hsa-211106 gene is used as a template, and primers are used to perform PCR amplification on piR-hsa-211106 to generate a double Chain PIWI protein-interacting RNA piR-hsa-211106, for the antisense strand of PIWI protein-interacting RNA piR-hsa-211106, carry out 3' end cholesterol modification, 3' end four sulfur skeleton modification, 5' end two sulfur Skeleton modification and full-chain methoxy modification (Shanghai Gemma Pharmaceutical Technology Co., Ltd.) completed the construction of the agonist.

Embodiment 2

[0033] This embodiment involves the process of transfecting the agonist into lung adenocarcinoma cells: the lung adenocarcinoma cells are subcultured until the cell density reaches about 30-50%, and the agonist is transfected into the lung adenocarcinoma cells by using lipo3000. Take 5 μl (100 pmol) of agonist and add it into the EP tube containing 95 μl DMEM medium, blow and mix well, then take 5 μl lipo3000 and add it into the EP tube containing 95 μl DMEM medium, blow and mix well, and mix the two at a volume of 1:1 The transfection mixture was formed and added to the culture plate after standing at room temperature for 20 minutes. Gene and protein levels were analyzed 48 hours later.

Embodiment 3

[0035] This example involves the analysis process of the proliferation activity of lung adenocarcinoma cells A549 and HCC2279 CCK8: lung adenocarcinoma cells were cultured, and the experimental group was transfected with PIWI protein-interacting RNA piR-hsa-211106. , 3 and 4 days later, the lung adenocarcinoma cells in the experimental group and the control group were aspirated and discarded the original medium, and the lung adenocarcinoma cells were washed twice with phosphate buffered saline (PBS), and 90 μl of fresh medium and 10 μl of CCK8 reagent ( Nanjing Nuoweizan Biotechnology Co., Ltd.) was treated for 1 hour, and the OD value was detected at the absorbance of 450nm. The OD value represented the cell density and reflected the cell proliferation activity. The results were as follows figure 1 As shown, the agonist constructed by PIWI protein-interacting RNA piR-hsa-211106 can significantly inhibit the proliferation of lung adenocarcinoma cells A549 and HCC2279.

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Abstract

The invention belongs to the technical field of biomedicine, and specifically relates to the use of PIWI protein interaction RNApiR-hsa-211106, which is used to prepare targeted therapeutic drugs for inhibiting the proliferation of lung adenocarcinoma cells, and is an abnormal activation of the cell energy metabolism pathway of lung adenocarcinoma PIWI protein-interacting RNA piR‑hsa‑211106 inhibits the content of pyruvate carboxylase and then inhibits the tricarboxylic acid cycle process, significantly inhibits energy metabolism and the proliferation and migration of lung adenocarcinoma cells ability, thereby promoting the apoptosis of lung adenocarcinoma cells, which directly acts on the target site without producing toxic side effects and off-target phenomena; through high-throughput sequencing analysis, mass spectrometry analysis technology and in vivo and in vitro experiments, it has been confirmed that PIWI protein interacts with RNA piR‑hsa‑211106 has strong specificity, high targeting rate, and less off-target effects and side effects.

Description

Technical field: [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the use of PIWI protein interaction RNA piR-hsa-211106, which is used for inhibiting the proliferation of lung adenocarcinoma cells and preparing targeted therapeutic drugs for lung adenocarcinoma. Background technique: [0002] Lung cancer is the most common malignant tumor. According to data from GLOBOCAN, there were 2.1 million new cases of lung cancer in the world in 2018, accounting for 11.6% of all new tumor cases (ranking first); 1.8 million deaths, accounting for 10% of all cancer deaths. 18.4% (No. 1). Lung cancer includes small cell lung cancer and non-small cell lung cancer. Among them, non-small cell lung cancer accounts for about 85% of all lung cancers, and lung adenocarcinoma is the most common histological type of non-small cell lung cancer, accounting for about 40-40% of primary lung tumors. 50%, most of which originated from the bronchial mucosa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K31/713A61P35/00C12N15/113
CPCA61K31/713A61P35/00C12N15/113C12N2310/10
Inventor 徐文华刘永梅董妍涵高金宁郝晓丹王子博李濛
Owner QINGDAO UNIV
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