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Application of liver cancer diagnosis and treatment oncogenic kinase marker

A technology of diagnostic markers and therapeutic drugs, applied in the field of biology

Active Publication Date: 2020-12-22
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

STK39 plays an important role in renal salt transport and blood pressure regulation by regulating the body's ion homeostasis, but there is no report on its role and mechanism in the occurrence and development of liver cancer

Method used

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  • Application of liver cancer diagnosis and treatment oncogenic kinase marker
  • Application of liver cancer diagnosis and treatment oncogenic kinase marker
  • Application of liver cancer diagnosis and treatment oncogenic kinase marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Detection of STK39 expression level in liver cancer tissue

[0062] main reagent

[0063] STK39 primers (GenScript), TRIzol (Tiangen), reverse transcription kit (ABMgood), SYBR (Yisheng), STK39 antibodies (abcam and sigma), immunohistochemical related reagents;

[0064] main instrument

[0065] PCR and QPCR instrument, Western Blot electrophoresis instrument, chemiluminescence imaging analysis system;

[0066] main method

[0067] QPCR

[0068] Human liver cancer tissue or cell samples were lysed with TRIzol, and RNA was extracted with chloroform and isopropanol, and then the RNA was reverse-transcribed into cDNA. 250 ng of cDNA was used as a template for QPCR quantitative detection, and GAPDH was used as an internal reference. The forward primer sequence of STK39 is 5′-TTCATAAAACCGAAGACGGG-3′, the reverse primer sequence is 5′-GTATTTGTTCGGGGATGGTG-3′; the forward sequence of GAPDH primer is 5′-ACCCAGAAGACTGTGGATGG-3′, and the reverse primer sequence is 5′-TTCAGCTCA...

Embodiment 2

[0076] Knockdown or deletion of STK39 can inhibit the proliferation of liver cancer cells

[0077] main reagent

[0078] STK39 antibody (abcam), trypan blue (Yisheng), CCK8 (Yisheng), crystal violet (Sangon Biotech);

[0079] main instrument

[0080] Cell counter, microplate reader;

[0081] main method

[0082] live cell count

[0083] The tumor cells were seeded in 12-well plates (50,000 / well), and the cells were digested on the 2nd, 3rd, 4th, and 5th day respectively, and the living cells were counted by trypan blue staining.

[0084] CCK8

[0085] Tumor cells were inoculated in a 96-well plate (2,000 / well), and CCK8 (10 μL) was added according to the time marked in the figure, and the value was read at 450 nm after 1.5 hours of reaction.

[0086] clone formation

[0087] Tumor cells were inoculated in 6-well plates (2,000 / well), and fresh medium was changed every 3 days. After 15 days, the cells were stained with crystal violet and photographed.

[0088] result

...

Embodiment 3

[0091] Knockdown or deletion of STK39 inhibits tumor formation in liver cancer

[0092] main reagent

[0093] Chloral hydrate;

[0094] main method

[0095] After nude mice were anesthetized for 4-6 weeks, tumors were subcutaneously mounted (2 million / mouse). The tumor growth size was counted every week, and the mice were sacrificed after 5 weeks, the tumor was stripped, the tumor weight was measured, and the tissue was fixed for pathological sectioning.

[0096] result

[0097] In order to further explore the important role of STK39 in the occurrence and development of liver cancer, the applicant of the present invention carried out tumor-bearing nude mice by knocking down and knocking out STK39 liver cancer cells. The results showed that knockdown or knockout of STK39 could significantly inhibit the formation ability of liver cancer tumors ( image 3 ), revealing that STK39 is essential for the growth of liver cancer.

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Abstract

The invention discloses an application of an oncogenic kinase marker for liver cancer diagnosis and treatment, and discloses an application of oncogenic kinase STK39 in liver cancer diagnosis and treatment. The expression difference of the STK39 in normal liver tissues and liver cancer tissues is found for the first time, the expression quantity of the STK39 in the liver cancer tissues is obviously higher than that in the normal tissues, and a patient with high expression of the STK39 has poor prognosis, so that the STK39 is prompted to be closely related to occurrence, development and prognosis of the liver cancer. Knockdown, knockout and inhibition of STK39 can obviously inhibit the growth and invasion of hepatoma carcinoma cells, and it is indicated that STK39 can be used as a new target for diagnosis and treatment of hepatoma carcinoma. A kit and the technology for checking STK39 expression can be applied to liver cancer diagnosis and prognosis prediction, and treatment strategiessuch as antibodies or small molecule inhibitors of targeted oncokinase STK39 are expected to be applied to liver cancer treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of an oncogenic kinase marker for the diagnosis and treatment of liver cancer. Background technique [0002] Liver cancer is a common malignant tumor with the characteristics of high incidence, low survival rate and extremely poor clinical treatment effect. The incidence of liver cancer ranks fifth among all malignant tumors, but its fatality rate ranks second among all malignant tumors. According to statistics, nearly 850,000 new liver cancer patients are diagnosed each year in the world, and about 750,000 people die of liver cancer. What is more noteworthy is that among all liver cancer patients, more than 50% of the patients are located in China, that is, China is a big country for liver cancer. In the face of such a huge threat of tumors, human beings have very few ways to deal with it. Although HBV, HCV infection, long-term alcohol consumption, a...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/573A61K45/00A61P35/00
CPCG01N33/57438G01N33/573A61K45/00A61P35/00G01N2800/52
Inventor 夏洪平张成飞周国仁许萍莫骁徐昊珺
Owner NANJING MEDICAL UNIV
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