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Molecular marker LncRNA DANCR for diagnosing and treating bladder cancer and application thereof

A molecular marker, bladder cancer technology, which can be used in medical preparations containing active ingredients, drug combinations, biochemical equipment and methods, etc., can solve the problem of not clearly elucidating the mechanism of lymphatic metastasis of bladder cancer.

Active Publication Date: 2019-02-22
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current research has revealed that lncRNA is involved in the biological functions of bladder cancer proliferation, metastasis, and apoptosis, it has not yet clearly elucidated the mechanism of lymphatic metastasis of bladder cancer.

Method used

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  • Molecular marker LncRNA DANCR for diagnosing and treating bladder cancer and application thereof
  • Molecular marker LncRNA DANCR for diagnosing and treating bladder cancer and application thereof
  • Molecular marker LncRNA DANCR for diagnosing and treating bladder cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 LncRNA DANCR is highly expressed in bladder cancer and is associated with lymphatic metastasis and poor prognosis

[0030] In this example, real-time fluorescent quantitative PCR and RNA fluorescence in situ hybridization were used to detect the expression of LncRNA DANCR in bladder cancer tissues.

[0031] (1) RNA extraction and reverse transcription-real-time fluorescent quantitative PCR experiment

[0032] 1. Total RNA extraction: ①Tissue RNA lysis: Grind the frozen fresh tissue of bladder cancer into small particles in liquid nitrogen, add 1ml Trizol lysate, gently pipette and mix to fully lyse the cells, and transfer the lysate to 1.5ml EP Tube, let stand at room temperature for 5 min. ② Lysis of cell RNA: Aspirate the medium, wash 2 times with PBS, every 10 6Add 1ml Trizol lysate to each cell, gently pipette and mix to fully lyse the cells, transfer the lysate to a 1.5ml EP tube, and let stand at room temperature for 5min. Add 1 / 5 volume of chloroform...

Embodiment 2

[0054] Example 2 Silencing LncRNA DANCR inhibits the proliferation ability of bladder cancer cells in vitro

[0055] In this example, siRNA1 (si-DANCR-1), siRNA2 (si-DANCR-2) and siRNA3 (si-DANCR-3) were used to silence LncRNA DANCR, and si-Ctrl was used as a control. The siRNA1, siRNA2, siRNA3 and si-Ctrl were synthesized by Shanghai Gemma. The siRNA1 sequence is shown in SEQ ID NO:5; the siRNA2 sequence is shown in SEQ ID NO:6; the siRNA3 sequence is shown in SEQ ID NO:7; the si-Ctrl sequence is shown in SEQ ID NO:8 shows:

[0056] SEQ ID NO: 5: 5'-GCGUACUAACUUGUAGCAA-3';

[0057] SEQ ID NO: 6: 5'-GAGCUAGAGCAGUGACAAU-3';

[0058] SEQ ID NO: 7: 5'-GUUGACAACUACAGGCACA-3';

[0059] SEQ ID NO: 8: 5'-CAACAAGAUGAAGAGCACC-3'.

[0060] 1. Bladder cancer cell culture

[0061] Bladder cancer cell lines UM-UC-3 and T24 were purchased from the American Type Culture Collection (ATCC). UM-UC-3 was cultured in DMEM medium, and T24 was cultured in RPMI-1640 medium. The medium contain...

Embodiment 3

[0075] Example 3 Silencing LncRNA DANCR inhibits the migration and invasion ability of bladder cancer cells in vitro

[0076] In this example, siRNA1 (si-DANCR-1), siRNA2 (si-DANCR-2) and siRNA3 (si-DANCR-3) were used to silence LncRNA DANCR, and si-Ctrl was used as a control. The siRNA1, siRNA2, siRNA3 and si-Ctrl are the same as in Example 2.

[0077] 1. Cell migration experiment

[0078] (1) The Transwell chamber was purchased from Corning Company in the United States, with a pore size of 8 μm. Before use, add 200 μl of medium containing 10% serum to the upper chamber to make the membrane hydrophilic, and absorb it before adding cells;

[0079] (2) Digest the cell culture 48h after transfection, resuspend the cells with medium containing 1% serum, count, and adjust the concentration to 4×10 5 / ml;

[0080] (3) Add 600 μl of medium containing 10% serum to the lower chamber, add 200 μl of cell suspension to the upper chamber, and continue culturing in the incubator for 13 ...

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Abstract

The invention discloses a molecular marker for diagnosing and treating the bladder cancer, namely LncRNA DANCR, and also discloses application of the molecular marker to screening or preparation of reagents used for diagnosing the bladder cancer diagnosing, bladder cancer lymphatic metastasis, bladder cancer prognosis relapse or bladder cancer clinical stages and application of the molecular marker to screening or preparation of drugs used for treating the bladder cancer. It is found that the LncRNA DANCR has high expression in bladder cancer cases with lymphatic metastasis, has negative correlation with overall survival prognosis of a patient and is an independent indicator for diagnosing the bladder cancer and judging bladder cancer development and survival prognosis; silencing of the LncRNA DANCR can restrain in-vitro proliferation, cell migration and invasion and in-vivo tumor formation and lymphatic metastasis of bladder cancer cells. It is verified that the LncRNA DANCR is an important cancerigenic factor of the bladder cancer for the first time, and can be used as a molecular marker for bladder cancer prognosis diagnosis and a new target spot for treatment.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis and biomedicine, and in particular relates to a molecular marker LncRNA DANCR for diagnosis and treatment of bladder cancer and its application. Background technique [0002] Bladder cancer (BCa) is the most common malignant tumor of the male genitourinary system in my country. According to the depth of tumor cell invasion, it is divided into non-muscle invasive bladder cancer (Ta-1) and muscle invasive bladder cancer (T2-4), the latter is prone to metastasis, progression and postoperative recurrence. The main cause of death from bladder cancer is metastasis, and lymphatic metastasis is the main and primary mode. Lymphatic metastasis is one of the most important factors affecting the prognosis of patients. Once lymphatic metastasis occurs, the 5-year survival rate is only 25-35%. Therefore, early diagnosis, accurate prediction and reasonable treatment of lymphatic metastasis of blad...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00A61P35/00C12Q1/6886C12Q2600/118C12Q2600/178
Inventor 陈旭谢锐辉林天歆陈子月
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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