Molecular marker HSF1 for diagnosing and treating bladder cancer and application thereof

A technology of molecular markers and bladder cancer, applied in medical preparations containing active ingredients, analytical materials, drug combinations, etc., can solve problems such as method limitations and poor therapeutic effects

Pending Publication Date: 2021-12-31
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

In addition, the current clinical treatment for patients with lymphatic metastasis of bladder cancer is mainly surgery combined with neoadjuvant chemotherapy or postoperative chemotherapy, which is limited and the treatment effect is not good. Reasonable treatment plan is extremely important to prolong the survival of patients

Method used

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  • Molecular marker HSF1 for diagnosing and treating bladder cancer and application thereof
  • Molecular marker HSF1 for diagnosing and treating bladder cancer and application thereof
  • Molecular marker HSF1 for diagnosing and treating bladder cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 HSF1 is highly expressed in bladder cancer, which is positively correlated with lymphatic metastasis and poor prognosis

[0022] In this example, real-time fluorescent quantitative PCR and immunohistochemical methods were used to detect the expression of HSF1 in bladder cancer tissues.

[0023] (1) RNA extraction, reverse transcription and real-time fluorescent quantitative PCR experiment

[0024] 1. Total RNA extraction: ①Tissue RNA lysis: Grind frozen fresh bladder cancer tissue into small particles in liquid nitrogen, add 1ml Trizol lysate, mix well to fully lyse the cells, and transfer to a 1.5ml EP tube. Stand at room temperature for 5 minutes. ② Lysis of cell RNA: Aspirate the medium, wash 2 times with PBS, every 10 6 Add 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 to the lysate, vortex vigorously to mix,...

Embodiment 2

[0048] Example 2 Silencing HSF1 inhibits the in vitro proliferation and in vivo tumorigenic ability of bladder cancer cells

[0049] In this example, SiRNA-1 (Si-HSF1-1), SiRNA-2 (Si-HSF1-2), SiRNA-3 (Si-HSF1-3) and SiRNA-4 (Si-HSF1-4) were used to silence HSF1 , while using Si-Ctrl as a control. The SiRNA and Si-Ctrl were synthesized by Shanghai Gemma Company. The siRNA-1 sequence is shown in SEQ ID NO: 3; the siRNA-2 sequence is shown in SEQ ID NO: 4; the siRNA-3 sequence is shown in SEQ ID NO: 5; the siRNA-4 sequence The sequence is shown in SEQ ID NO:6; the sequence of the Si-Ctrl is shown in SEQ ID NO:7:

[0050] SEQ ID NO: 3: 5'-UAGCCUGCCUGGACAAGAA-3';

[0051] SEQ ID NO: 4: 5'-GUGCUGCCCAAGUACUUCA-3';

[0052] SEQ ID NO: 5: 5'-GAGUGAAGACAUAAAGAUC-3';

[0053] SEQ ID NO: 6: 5'-GCGGCAGCUCAACAUGUAU-3';

[0054] SEQ ID NO: 7: 5'-UUCUCCGAACGUGUCACGU-3';

[0055] (1) Cell culture and transfection

[0056] 1. Bladder cancer cell culture

[0057] Bladder cancer cell lin...

Embodiment 3

[0076] Example 3 Silencing HSF1 inhibits bladder cancer cell migration and invasion in vitro and lymphatic metastasis in vivo

[0077] In this example, SiRNA-1 (Si-HSF1-1) and SiRNA-2 (Si-HSF1-2) were used to silence HSF1 respectively, and Si-Ctrl was used as a control. The SiRNA-1, SiRNA-2 and Si-Ctrl are the same as in Example 2.

[0078] (1) Migration and invasion experiments of bladder cancer cells in vitro

[0079] 1. Cell migration experiment

[0080] (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 serum-free medium to the upper chamber to make the membrane hydrophilic, and absorb it before adding cells;

[0081](2) Digest the cell culture 48h after transfection, resuspend the cells with serum-free medium, count, and adjust the concentration to 2×10 5 / ml medium;

[0082] (3) Add 700 μl of medium containing 10% serum to the lower chamber, add 200 μl of cell suspension to the upper ...

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PUM

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Abstract

The invention provides a molecular marker HSF1 for diagnosing and treating bladder cancer and an application thereof. Comparison of gene expression results in a bladder cancer clinical specimen shows that HSF1 is highly expressed in bladder cancer tissue with positive lymphatic metastasis, and negatively correlated with overall survival prognosis. The molecular marker can be used as a marker to indicate lymphatic metastasis in bladder cancer, and as an independent indicator to predict survival prognosis of a patient. In-vivo and in-vitro function experiments of bladder cancer cells show the silencing HSF1 can inhibit proliferation of bladder cancer cells and lymphatic metastasis. The invention discovers that HSF1 is an important carcinogenic factor of bladder cancer for the first time, and the HSF1 can be used as a molecular marker for diagnosing bladder cancer and judging prognosis, and as a new target for treating bladder cancer.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis and biomedicine, and specifically relates to a molecular marker HSF1 for diagnosing and treating bladder cancer and its application. Background technique [0002] In 2020, there will be 573,278 new cases of bladder cancer worldwide, with 212,536 deaths, ranking 10th among all malignant tumors and the most common malignant tumor of the urinary system in China. It is worth noting that about 25% of bladder cancer patients have muscle-invasive bladder cancer, which has a higher risk of lymphatic metastasis. Compared with patients with negative lymphatic metastasis, the five-year survival rate of patients with positive lymphatic metastasis will be reduced by 70%. % dropped sharply to 25%-31%, and the postoperative recurrence rate was more than 3 times that of patients with negative lymphatic metastasis. In addition, the current clinical treatment for patients with lymphatic metastasis of b...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574A61K31/7105A61P35/00
CPCC12Q1/6886G01N33/57407G01N33/57484A61K31/7105A61P35/00C12Q2600/118C12Q2600/158Y02A50/30
Inventor 林天歆陈旭黄铭董文
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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