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Sorafenib drug-resistant marker and application thereof

A fenib-resistant and marker technology, applied in the field of biomedicine, can solve problems such as disease progression

Pending Publication Date: 2021-12-17
THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high degree of heterogeneity within and between individuals in liver cancer, a considerable number of patients do not respond to sorafenib treatment, and even if there are responding patients, the treatment response is often at most about 4 months (median PFS and TTP Both were 3.7 months), followed by drug resistance, and disease progression after drug resistance

Method used

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  • Sorafenib drug-resistant marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The expression level of CCT6A in liver cancer tissues and its relationship with the prognosis of liver cancer patients.

[0050] 1. Method:

[0051] 1.1 Using tumor gene database ONCOMINE (https: / / www.oncomine.org / resource / login.html), UALCAN (http: / / ualcan.path.uab.edu / ) and GEPIA (http: / / gepia.cancer- pku.cn / ) to analyze the expression of various genes and proteins in liver cancer.

[0052] 1.2 Using human liver cancer tissue samples to detect the level and distribution of CCT6A in tissues

[0053] (1) Collect 200 cases of liver cancer after surgical resection and paired paracancerous tissues, extract total RNA and total protein, and prepare tissue sections; collect various clinical data, follow-up data, etc.

[0054] (2) Immunohistochemical method:

[0055] The experimental steps are as follows: first set the oven at 60°C for 30 minutes to dewax the slices; then soak the wax in xylene, 2 cylinders, 10 minutes each time, 2 times in total. Next hydration: 100% alco...

Embodiment 2

[0087] Co-immunoprecipitation and co-localization studies of CCT6A and B55.

[0088] 1. Method:

[0089] 1.1 In order to further explore the potential regulatory factors of CCT6A affecting the progression of liver cancer, we predicted the protein network interacting with CCT6A through STRING and the database (https: / / string-db.org / ) and GEPIA database respectively.

[0090] 1.2 Construct CCT6A plasmid with Flag tag and B55γ plasmid with Ha tag, transfect liver cancer cell HepG2 through exogenous expression system, and use Western blotting to detect the expression of the tag.

[0091] 1.3 The plasmid constructed in 1.2 above was co-transfected into HepG2 cell line, and the cells were collected 48 hours later. The combination of CCT6A and B55γ was detected by immunoprecipitation technique, and the co-localization of CCT6A and B55γ in the cytoplasm was detected by immunofluorescence. Using the UniProt (https: / / www.uniprot.org / ) database to analyze the protein structure of CCT6A ...

Embodiment 3

[0107] CCT6A knockdown promotes autophagy in hepatoma cells and provides sorafenib sensitivity studies.

[0108] 1. Method

[0109] 1.1 Construction of CCT6A knockdown liver cancer cell lines

[0110] CCT6A knockdown liver cancer cell lines (MHCC-97L and HepG2) were constructed by lentiviral transfection system, nonsense sequence and empty vector were used as negative controls, and CCT6A expression changes were detected by Western blotting for verification.

[0111] Lentiviral infection expression construction method:

[0112] The pLKO.1 vector (Sigma-Aldrich, USA) was double-digested with restriction endonucleases ageI and EcoRI, and the digested products were recovered with low-melting point agarose gel and mixed with the annealed products of the above oligonucleotides. Ligation was then performed overnight with T4 DNA ligase. The shCCT6A sequence is:

[0113] shCCT6A-1# (MHCC-97L), 5'-CCGGCCAGACATCTCTTCGTACTACTCGAGTAGTACAGAGATGTTCTGGTTTTTG-3' (Forward) (SEQ ID NO.3);

...

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Abstract

The invention relates to a sorafenib drug-resistant marker and application thereof, and belongs to the technical field of biomedicine. The sorafenib drug-resistant marker is CCT6A. Research of the marker finds that on one hand, the relationship between CCT6A / B55 gamma pathway regulation and control of autophagy and liver cancer cell sorafenib drug resistance can be identified through deep research, and a scientific basis is provided for a new approach of targeting the pathway to treat liver caner. On the other hand, sorafenib drug-resistant people can be prompted, and the effects of personalized treatment and precise treatment are achieved in liver cancer treatment or cancer treatment.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a Sorafenib resistance marker and its application. Background technique [0002] Liver cancer is one of the deadliest cancers and the third most common cause of cancer death worldwide. According to the latest global cancer data released by the World Health Organization, there will be 910,000 new cases of liver cancer and 830,000 deaths in 2020, showing a clear upward trend and seriously endangering human health. Due to the insidious onset of liver cancer and the insufficient promotion of general screening or screening, most patients are diagnosed at an advanced stage. Currently, the targeted drug Sorafenib is the drug of choice for advanced liver cancer. Sorafenib is an oral multikinase inhibitor. It can inhibit tumor cell proliferation and angiogenesis, and promote tumor cell apoptosis. It was approved as the first targeted drug for the treatment of advanced HCC in 2007. H...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574
CPCC12Q1/6886G01N33/57484G01N33/57438C12Q2600/106C12Q2600/118C12Q2600/158
Inventor 黄月华练一帆黄彦霖曾国芬王嘉亮
Owner THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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