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Application of HSDL1 to diagnosis and treatment of osteosarcoma

A technology for osteosarcoma and products, applied in the application field of HSDL1 in the diagnosis and treatment of osteosarcoma, can solve the problems of metastasis and recurrence rate, and achieve the effect of early diagnosis, more accurate gene diagnosis and timely gene diagnosis

Active Publication Date: 2017-05-24
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although chemotherapy has played an important role in the treatment of malignant osteosarcoma since the emergence of chemotherapy in the last century, the control of its metastasis and recurrence rate is still unsatisfactory.

Method used

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  • Application of HSDL1 to diagnosis and treatment of osteosarcoma
  • Application of HSDL1 to diagnosis and treatment of osteosarcoma
  • Application of HSDL1 to diagnosis and treatment of osteosarcoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 High-throughput sequencing and analysis

[0055] The tumor tissues and adjacent tissues of osteosarcoma patients were selected for gene comparison, and the tumor tissues and adjacent tissues of 6 osteosarcoma patients were selected. All tissue samples and pathological results should be true and reliable. The research was approved by the ethics committee and the patients were informed. agree.

[0056] RNA extraction is performed on tumor tissue and para-cancerous tissues, and after RNA extraction, agarose gel electrophoresis can be used to preliminarily determine whether the quality of the extracted RNA sample is qualified or not, and whether it can be used for further transcriptome analysis. Then, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the sample requirements for RNA-seq sequencing: OD260 / OD280 was 1.8-2.2.

[0057] The sequencing platform is the HiSeq 2500 high-throughput sequencing platform of Illumina, which perfo...

Embodiment 2

[0058] Example 2 Differential expression of HSDL1 gene in paracancerous tissue and tumor tissue

[0059] 1. Sample acquisition: 40 patients with osteosarcoma, the paracancerous tissues and tumor tissues were collected respectively.

[0060] 2. Acquisition of RNA in samples

[0061] The RNA extraction kit of QINGEN Company was used to extract the total RNA of cells according to the instruction.

[0062] 3. Reverse transcription

[0063] The reverse transcription of RNA was carried out using the reverse transcription kit of TAKARA company.

[0064] 4. QPCR

[0065] (1) Primer design

[0066] QPCR amplification primers were designed according to the coding sequences of HSDL1 gene and GAPDH gene in Genbank, and synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. The specific primer sequences are as follows:

[0067] HSDL1 gene:

[0068] The forward primer is 5'-AAGGACCACAGGATATTG-3' (SEQ ID NO.3);

[0069] The reverse primer is 5'-AAGGACCACAGGATATTG-...

Embodiment 3

[0082] Example 3 Differential expression of HSDL1 protein

[0083] 1. The research object is the same as in Example 2.

[0084] 2. Extract tissue total protein

[0085] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0086] 3. Western blot detection

[0087] The extracted protein was quantitatively subjected to SDS-PAGE electrophoresis, followed by membrane transfer, blocking, primary antibody incubation, secondary antibody incubation, and color development.

[0088] 4. Statistical processing

[0089] The gray value of the protein band was analyzed using Image J software, and the gray value of the HSDL1 protein band was normalized with β-actin as an internal reference. The results and data are expressed in the form of mean ± standard deviation, and SPSS13.0 statistical software is used for statistical analysis. The difference between the two is tested by t test, and it is considered statistically significant when P<...

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Abstract

The invention discloses application of HSDL1 to diagnosis and treatment of osteosarcoma and particularly relates to an HSDL1 gene and an expression product thereof to application of a diagnosis and treatment tool of osteosarcoma. A high-throughput sequencing and RT-PCR (Reverse Transcription-Polymerase Chain Reaction) experiment proves that the expression of the HSDL1 gene in pericarcinomatous tissues and the expression of the HSDL1 gene in tumor tissues have difference; and the expression of the HSDL1 gene in the tumor tissues is remarkably reduced. Therefore, the HSDL1 gene can be used as a molecular marker for treating and diagnosing osteosarcoma. Furthermore, enlarged sample detection shows that the HSDL1 has very good relevance with osteosarcoma and can be used for preparing an osteosarcoma assisted diagnosis preparation. The aim of promoting gene expression can be realized through constructing an over-expression vector of the gene, so that the aim of treating cancers is realized.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the technical field of early diagnosis and treatment of osteosarcoma, and in particular to the application of HSDL1 in the diagnosis and treatment of osteosarcoma. Background technique [0002] Osteosarcoma (is one of the most common malignant primary tumors of bone (except for installed cell tumors), originating from mesenchymal tissue, the main feature is mainly spindle-shaped stromal cells that can produce bone-like tissue. In terms of clinical characteristics Osteosarcoma tends to occur in adolescents, with hidden symptoms at onset, atypical symptoms, and high degree of malignancy. In the early stage of disease discovery, distant metastasis often occurs, and the rate of lung metastasis is high. Therefore, osteosarcoma is very harmful and fatal. The high rate seriously affects the health and life of young patients. Although chemotherapy has played an important role in the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574G01N33/573
CPCC12Q1/6886G01N33/573G01N33/57407G01N2333/902
Inventor 孙耀兰任静边洋
Owner GUAN BOJIAN BIOTECH CO LTD
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