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Method for detecting anti-liver cancer efficacy of tyroserleutide, kit and gene chip used thereby

A gene chip, anti-cancer technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., can solve problems such as unpredictability of patients

Inactive Publication Date: 2006-09-27
SHENZHEN KANGZHE PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it often takes a long period of time from the administration of the drug to the change in the shape of the tumor. During this period, it is unpredictable whether the drug will be effective for the patient.

Method used

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  • Method for detecting anti-liver cancer efficacy of tyroserleutide, kit and gene chip used thereby
  • Method for detecting anti-liver cancer efficacy of tyroserleutide, kit and gene chip used thereby
  • Method for detecting anti-liver cancer efficacy of tyroserleutide, kit and gene chip used thereby

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0063] YSL anti-human liver cancer BEL-7402 xenograft tumor experiment in nude mice

[0064] For the preparation of human liver cancer model in nude mice, please refer to the literature (Han Rui. Anticancer Drug Research and Experimental Technology [M]. First Edition. Beijing: Beijing Medical University and China Union Medical University Press. 1997, 4: 299).

[0065] BEL-7402 nude mice bearing human liver cancer were purchased from the Animal Center of Cancer Institute, Chinese Academy of Medical Sciences. They were reared in a laminar flow cabinet with independent ventilation without specific pathogens, and the feed, litter and drinking water used were all aseptically treated in strict accordance with the NIH standard (safety operation number: A3873-1).

[0066] In the present invention, select nude mice bearing human liver cancer BEL-7402 with a tumor diameter greater than 1 cm and a good growth state, and cut the fresh tumor tissue into 2-4 mm under sterile conditions. 3 ...

Embodiment 1

[0068] Reverse transcription PCR (RT-PCR) detection of YSL on mitochondrial respiratory chain-related genes NDUFA2, NDUFA10, NDUFS1, SUCLG1, calcium homeostasis regulatory protein gene Calreticulin, and genes PTEN, Akt1 / 2 in BEL-7402 nude mouse xenograft tumor cells , p27, p21 mRNA expression

[0069] Specific steps are as follows:

[0070] (1) Sample preparation

[0071] In the YSL anti-human liver cancer BEL-7402 nude mouse tumor transplantation experiment, 5 tumor-bearing nude mice were randomly selected from each group from the normal saline group and the tyroserleutide 160 μg / kg / d dose group, and were killed by cervical dislocation. Remove the tumor tissue with RNase-free ophthalmic scissors, select a part with good tumor growth, no obvious necrosis, ulceration, and hard texture, cut it into small pieces of about 50 mg, wrap it in tin foil, and immediately put it into liquid nitrogen for freezing spare.

[0072] (2) Extraction of total RNA

[0073] 1) Tissue homogenate:...

Embodiment 2

[0091] Kit for detecting the effect of tyroserleutide

[0092] The kit has a box body, and the box body includes sealed containers respectively filled with solid dry powder primers of SEQ ID Nos. 1-22.

[0093] The kit may also contain sealed containers respectively containing the following substances:

[0094] DEPC-treated sterilized double distilled water, 2.5mmol / L dNTP, 15mmol / L magnesium chloride, 10×PCR buffer, AMV reverse transcriptase, RNase inhibitor, Taq DNA polymerase.

[0095] Wherein, the preparation (100ml volume) of 10 * PCR buffer solution is as follows:

[0096] DEPC treated sterile double distilled water 70mL

[0097] Tris(Tris) 0.158g

[0098] Potassium chloride 0.373g

[0099] Triton X-100 0.1mL

[0100] Hydrochloric acid Adjust pH to 9.0

[0101] DEPC-treated sterilized double-distilled water was added to 100mL

[0102] When the kit is used, the solid dry powder primers and sterilized double-distilled water can be first prepared into a solution at a...

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Abstract

The invention discloses a detecting method and its agent box and gene chip of tyr-ser-leu protein anti-hepatocarcinoma effect, which comprises the following steps: detecting the changing condidtions of gene NDUFA2, NDUFA10, NDUFS1, SUCLG1, Calreticulin, PTEN, Akt1, Akt2, P21and P27 expression level in the biological material after acted by tyr-ser-leu protein; displaying downward for gene NDUFA2, NDUFA10,NDUFS1,SUCLG1,Akt1,Akt2 and upward for the expression level of gene Calreticulin,PTEN, P21,P27; possessing anti-hepatocarcinoma effect. The invention provides a rapid detecting path of tyr-ser-leu protein anti-hepatocarcinoma effect on the molecular level.

Description

technical field [0001] The invention relates to a method and a preparation for detecting the anticancer effect of drugs, in particular to a method for detecting the anticancer effect of tyroserleutide at the molecular level, a kit and a gene chip used therefor. Background technique [0002] Primary liver cancer is one of the most common malignant tumors of the digestive tract in humans. Its incidence rate ranks eighth among major cancers in the world, and its mortality rate is high. It is second only to gastric cancer and esophageal cancer in the mortality rate of malignant tumors and ranks third. . China is a country with a high incidence of primary liver cancer. Its morbidity and mortality rates rank first in the world, and its mortality rate ranks second among various cancers. About 110,000 people die from liver cancer every year, accounting for about 100,000 deaths from liver cancer in the world. about 45%. Currently, surgery, radiotherapy and chemotherapy are the main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 姚智陆融
Owner SHENZHEN KANGZHE PHARM CO LTD
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