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PNLIPRP3 gene and application of expression product of gene in tongue squamous carcinoma diagnosis and treatment

A technology for gene expression and tongue squamous cell carcinoma, which is applied in the field of prognosis prediction and tumor diagnosis, and can solve problems affecting the quality of life of patients, large lesion area, and wide range of surgical resection, etc.

Active Publication Date: 2017-06-13
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the size of the lesion is large and the scope of surgical resection is wide, which will seriously affect the quality of life of patients after surgery.

Method used

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  • PNLIPRP3 gene and application of expression product of gene in tongue squamous carcinoma diagnosis and treatment
  • PNLIPRP3 gene and application of expression product of gene in tongue squamous carcinoma diagnosis and treatment
  • PNLIPRP3 gene and application of expression product of gene in tongue squamous carcinoma diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening of differentially expressed genes

[0063] 1. Experimental materials

[0064] 5 tissue samples of tongue squamous cell carcinoma were obtained from oral and maxillofacial surgery patients, including 2 cases of well-differentiated squamous cell carcinoma, 2 cases of moderately differentiated squamous cell carcinoma, and 1 case of poorly differentiated squamous cell carcinoma; including 2 males and 3 females. At the same time, each case of cancerous tissue was selected as the normal tissue at >5cm around the cancerous tumor as its own control. All patients had not undergone chemotherapy, radiotherapy, biological therapy and other treatments for tumors before seeing the doctor. A part of the tissue was immediately stored in liquid nitrogen after collection.

[0065] 2. RNA extraction and cDNA synthesis

[0066] Total RNA was extracted with Trizol RNA reagent (Invitrogen Company), and total RNA was identified by UV spectrophotometer (ND-1000, NanoDrop...

Embodiment 2

[0075] Example 2 Verification of Differentially Expressed Genes in Large Samples

[0076] Based on the results of the primary screening of the microarray, we selected the PNLIPRP3 gene for large sample verification.

[0077] 1. Sample collection

[0078] According to the method of Example 1, 45 tongue squamous cell carcinoma tissues and 45 normal tissues were collected.

[0079] 2. Validation at the mRNA level

[0080] 2.1 Extract tissue RNA

[0081] Step is with embodiment 1.

[0082] 2.2 Reverse transcription

[0083] Reverse transcription using Primescript 1 st strand cDNA synthesis kit kit, the operation steps are as follows:

[0084] (1) Add the following reaction liquid in the microcentrifuge tube, as shown in Table 1:

[0085] Table 1 Reaction liquid

[0086] Reagent dose RNA 2.0μg dNTP 1.0μl Oligo(dT) 2.0μl RNase free dH 2 o

Add to 10.0μl

[0087] (2) Incubate at 70°C for 5 minutes, then rapidly cool to 4°C;

...

Embodiment 3

[0109] Example 3 PNLIPRP3 Gene Overexpression

[0110] 1. Construction of PNLIPRP3 gene recombinant plasmid

[0111] (1) amplifying the coding sequence of the PNLIPRP3 gene;

[0112] (2) Design amplification primers;

[0113] (3) The amplified PNLIPRP3 gene was connected to the expression vector pcDNA3.0 to construct the pcDNA3.0-PNLIPRP3 recombinant expression vector.

[0114] 2. Culture and transfection of tongue squamous cell carcinoma cells

[0115] 2.1 Cell culture

[0116] Human tongue squamous carcinoma cell line HN4 was cultured in DMEM / F12 medium containing 10% FBS, 100 U / m L penicillin and 100 μg / m L streptomycin. All cells were placed in 5% CO 2 in a 37 °C cell culture incubator.

[0117] 2.2 Cell transfection

[0118] (1) 24h before transfection, the cells were inoculated with 2×10 5 Inoculate each well in a 6-well plate, add DMEM / F12 medium, adhere to the wall overnight, and perform transfection when the cells reach 80-90% confluency.

[0119] (2) Dilute ...

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Abstract

The invention discloses a PNLIPRP3 gene which can be adopted as a molecular marker for tongue squamous carcinoma diagnosis and treatment. Experiments show that compared with normal tissue, the PNLIPRP3 gene is low in expression quantity in tongue squamous carcinoma tissue. The invention further discloses application of the PNLIPRP3 gene for preparing a drug for tongue squamous carcinoma treatment. The invention provides a novel tongue squamous carcinoma clinic treatment method, and further provides a novel drug target for tongue squamous carcinoma treatment.

Description

technical field [0001] The present invention relates to the fields of tumor diagnosis, treatment, and prognosis prediction. More specifically, the present invention relates to a method for tumor diagnosis and prognosis by detecting PNLIPRP3 abnormality; and a tumor therapeutic agent for activating PNLIPRP3 gene or protein. Background technique [0002] Oral squamous cell carcinoma is the most common malignant tumor of the head and neck, accounting for about 3% of systemic tumors and 90% of oral tumors, and tongue cancer ranks first in the incidence of oral squamous cell carcinoma. Although the incidence of oral cancer in developed countries has shown a slow downward trend in recent years, the overall incidence is increasing. Most of the patients are men over 40 years old, and the predilection sites are mostly on the tongue, cheeks, and gums. The occurrence of tongue squamous cell carcinoma is a multi-factor, multi-step, and multi-stage complex process, and the etiology is a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/156G01N33/57407G01N33/68
Inventor 马翠常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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