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Molecular marker related to incidence and development of laryngeal carcinoma

A technology of laryngeal squamous cell carcinoma and drugs, applied in the field of biomedicine, can solve the problems of high recurrence rate after surgery and difficulty in early diagnosis of laryngeal cancer

Active Publication Date: 2020-10-02
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reasons are mainly manifested in three aspects: First, the early clinical manifestations of laryngeal cancer are diverse, which brings great difficulty to the early diagnosis of laryngeal cancer. Most patients fail to receive timely and effective treatment in the early stage. Molecular markers are the key to improving the early diagnosis rate of laryngeal cancer
Second, the treatment of laryngeal cancer is mainly based on surgery, supplemented by radiotherapy and chemotherapy. Although partial laryngectomy and the early stage of glottic laryngeal cancer are treated with C02 laser treatment, the laryngeal function is completely or partially preserved, and postoperative complications Therefore, the quality of life of patients is improved, but the postoperative recurrence rate is still high. If the tumor-promoting genes or tumor suppressor factors of laryngeal cancer can be found out, and potential drug targets can be screened out, it will be a great opportunity for the development of laryngeal cancer. Cancer-related specific vaccines or / and new anti-cancer drugs lay the foundation

Method used

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  • Molecular marker related to incidence and development of laryngeal carcinoma
  • Molecular marker related to incidence and development of laryngeal carcinoma
  • Molecular marker related to incidence and development of laryngeal carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Molecular verification of differential expression

[0049] 1. Research object

[0050] The cancer tissues and corresponding paracancerous tissues of 45 patients with laryngeal squamous cell carcinoma were collected. Inclusion and exclusion criteria: ①Patients have no primary tumor other than the primary tumor, and the primary tumor has not metastasized; ②No history of cardiovascular and cerebrovascular diseases such as diabetes and hypertension; ③No infectious diseases such as hepatitis, syphilis, tuberculosis, and HIV Medical history; ④ no family history of genetic diseases; ⑤ no radiotherapy, chemotherapy and biological therapy before operation.

[0051] Sampling precautions: When cutting tumor specimens, pay attention to cutting non-necrotic tumor tissue in the central area of ​​the tumor. When cutting normal adjacent tissues, try to stay away from the tumor as far as possible, and be more than 1.5cm away from the edge of the tumor. Try to cut mucosal epit...

Embodiment 2

[0103] Example 2 LncRNA silencing

[0104] 1. Cell culture

[0105] The human laryngeal squamous carcinoma cell line Hep2 was incubated in RPMI1640 medium containing 10% fetal bovine serum and 1% P / S at 37°C and 5% CO 2 , cultured in an incubator with a relative humidity of 90%. The medium was changed once every 2-3 days, and the cells grew well and showed a monolayer adherent growth. Use 0.25% EDTA-containing trypsin for routine digestion and passaging.

[0106] 2. Transfection

[0107] 1) Treatment of cells before transfection

[0108] The day before transfection, 3~5×10 5 Cells / well were cultured in antibiotic-free medium for one day, and the cell density was 30-50% at the time of transfection, and replaced with serum-free medium before transfection.

[0109] 2) Design of siRNA

[0110] Two groups were set up in the experiment: negative control group (siNC, transfected with negative control siRNA), and siLncRNA silencing group (transfected with siRNA against LncRNA)....

Embodiment 3

[0131] Example 3 The effect of LncRNA expression on the proliferation of laryngeal squamous cell carcinoma cells

[0132] WST-1 method was used to detect the effect of lncRNA expression on the proliferation of laryngeal squamous cell carcinoma cells.

[0133] The next day after cell transfection, adjust the cell density to 1×10 2 / μl, seeded in a 96-well plate (100 μl / well), and each group set up three replicate wells. 37°C, 5% CO 2 After culturing for 72 hours, add 10 μl of WST-1 and incubate for 2 hours, then detect the optical density (D) value of the cells at a wavelength of 450 nm with a microplate reader.

[0134] The result is as figure 1 As shown, the difference between the groups was statistically significant (* P P <0.05). It shows that the expression of LOC730101, LOC442028, LOC100507437 and LOC158435 can inhibit the proliferation of laryngeal squamous cell carcinoma cells.

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Abstract

The invention discloses a molecular marker related to incidence and development of laryngeal carcinoma. The molecular marker is LOC100507437. According to the molecular marker related to incidence anddevelopment of laryngeal carcinoma, QPCR research proves that expression of LOC100507437 in laryngeal squamous cell carcinoma tissue is significantly up-regulated compared with those of control groups, and according to the differential expression characteristic of LOC100507437, LOC100507437 can be used as the molecular marker for diagnosing laryngeal squamous cell carcinoma. Moreover, an in-vitroexperiment proves that expression of LOC100507437 is related to proliferation and migration of laryngeal squamous cell carcinoma so that LOC100507437 can be used as a molecular target for treating laryngeal squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a molecular marker used for occurrence and development of laryngeal cancer. Background technique [0002] Laryngeal cancer is a common malignant tumor in the head and neck, accounting for the second place in the head and neck tumors. Its incidence rate accounts for about 5% of all malignant tumors in the whole body. Most squamous cell carcinomas originate in the vocal cords, accounting for about 60%. Huang Xinhui et al conducted a retrospective analysis on 306 cases of laryngeal malignant tumors and found that squamous cell carcinoma accounted for 99.7% of all malignant tumors, among which squamous cell carcinoma with high and moderate differentiation was the main type; the primary site of laryngeal cancer was glottic type The majority, followed by supraglottic type, subglottic type; smoking, drinking are closely related to the occurrence of tumors. Although surgery, radiotherapy, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K31/7088A61P35/00
CPCA61K31/7088A61K45/00A61P35/00C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 杨承刚孙耀兰
Owner BEIJING MEDINTELL BIOMED CO LTD
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