CN106841131A - A kind of detection method of biomarker based on keratin autofluorescence as predicted detection tumour and its application - Google Patents
A kind of detection method of biomarker based on keratin autofluorescence as predicted detection tumour and its application Download PDFInfo
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- CN106841131A CN106841131A CN201611243592.5A CN201611243592A CN106841131A CN 106841131 A CN106841131 A CN 106841131A CN 201611243592 A CN201611243592 A CN 201611243592A CN 106841131 A CN106841131 A CN 106841131A
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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Abstract
A kind of detection method of biomarker based on keratin autofluorescence as predicted detection tumour and application, step are as follows：After the tumor tissues of experimental subjects are subject to the light source irradiation comprising doses ultraviolet light within 72 hours, autofluorescence of the experimental subjects by keratin in the tumor tissues of the light source irradiation containing ultraviolet light is detected, wherein the dosage=uv power × irradiation time；Detect the spontaneous fluorescence intensity of the wavelength that is sent of tumor tissues that the experimental subjects irradiated by the light source containing ultraviolet light in the range of 400 640nm；The tumor tissues autofluorescence and the autofluorescence intensity of the tumor region tissue around irradiated by the light source containing ultraviolet light that the experimental subjects is irradiated by the light source containing ultraviolet light compare, and obtain the spontaneous fluorescence intensity rate of change induced through ultraviolet light；According to the keratin autofluorescence change rate of strength induced through ultraviolet light, the malignant tumor tissue position by experimental subjects is predicted.
Description
Technical field
Biomarker the present invention relates to predicting and detecting tumor tissues, specifically based on angle egg in detection tumor tissues
White autofluorescence change, as the detection method and its application of prediction and the biomarker of detection knub position.
Background technology
In epithelial cell, according to different cell categories, differentiation degree and functional status can all have special keratin
Expression.It turns out that, in tumour, epithelial tumor cell largely remains the hypotype of original keratin.According to this
Individual keratin is already used to as the mark that tumour cell is originated is differentiated, most common of which is gland cancer.
The method that it is critical only that tumor screening of present whole world early detection cancer.Current existing inspection method includes body
Various physiochemical indices are examined, B ultrasonic, X-ray, anal orifice and rectal intestine refer to inspection, the Pap smear, mammary gland molybdenum palladium in gynaecology's physical examination take the photograph piece etc..Simultaneously
Clinically, how to judge whether tumor tissues are thoroughly cut off rapidly in surgical procedure, also have great clinical meaning.
Section statining mainly is used in clinically performing the operation now, is judged that tumor tissues hand is no and is removed completely.This method
Not only need to wait for a long time, and can not completely determine whether that tumor tissues are cut off completely.
Studies have found that, Keratin 1 content is dramatically increased in some endothelium malignant tumor tissues.And so far without noninvasive inspection
The method for surveying Keratin 1 content in tissue.
So, it is badly in need of a kind of method of real-time detection tumor tissues.
The content of the invention
The purpose of the present invention is to overcome the deficiencies in the prior art, and the present invention can induce Keratin 1 autofluorescence based on ultraviolet
Rising this brand-new discovery, breach the problem of prior art, have found one kind can quickly detect exactly it is pernicious swollen
The biomarker of tumor tissue.And based on the biomarker, invent the method for new Non-invasive detection tumor tissues.
Concrete technical scheme of the invention is：
A kind of detection method of biomarker based on keratin autofluorescence as predicted detection tumour, comprises the following steps：
(1) after the tumor tissues in experimental subjects are subject to the light source comprising doses ultraviolet light to irradiate within 24 hours, detection
The experimental subjects passes through the autofluorescence of keratin in the tumor tissues that the light source containing ultraviolet light irradiates, wherein the dosage=
Uv power × irradiation time；
(2) detect wavelength that the experimental subjects sent by the tumor tissues that the light source containing ultraviolet light irradiates in 490-
Spontaneous fluorescence intensity in the range of 640nm；
(3) according to the method for the step (2), equally detect that the experimental subjects is subject to the swollen of the light source irradiation containing ultraviolet light
The autofluorescence intensity of knurl areas adjacent tissue；
(4) by the experimental subjects by the tumor tissues autofluorescence of the light source irradiation containing ultraviolet light and by containing ultraviolet light
The autofluorescence intensity of the tumor region tissue around of light source irradiation compares, and obtains strong through the autofluorescence of ultraviolet light induction
Degree rate of change；
(5) according to the keratin autofluorescence change rate of strength induced through ultraviolet light, it is right to be tested described in predicted detection
The malignant tumor tissue position of elephant.
Further, the tumour at the tumor tissues position such as including lung, liver, kidney, prostate, uterus, skin.
Further, the tumor type is gland cancer and cutaneum carcinoma.
Further, the tumour at the tumor tissues position such as including lung, liver, kidney, prostate, uterus, skin.
Further, in the detection sample in the method for Keratin 1 autofluorescence, by the ultraviolet polished bard of doses
Sharp effect is to amplify the signal of Keratin 1 autofluorescence.Uv agent amount is less than or equal between 100 joules/square centimeter.
Further, the mode of the utilization excitation Keratin 1 autofluorescence includes defeated using common continuous light
Go out excited mode, be modulated using electrical modulation in the mode that excites or the mode excited using pulse laser
At least one.
Further, the Keratin 1 autofluorescence is characterised by spectral region in 400-600nm.
Further, the Keratin 1 autofluorescence is characterised by spectral region in 450-550nm.
Further, the Keratin 1 autofluorescence is characterised by spectral region in 460-500nm.
The present invention also provides a kind of biomarker that malignant tumor tissue is detected based on detection Keratin 1 autofluorescence variation prediction
Application.
The present invention is significantly higher than normal structure this phenomenon using Keratin 1 autofluorescence intensity in malignant tumor tissue, is clinical
Detection malignant tumor tissue sets up basis, can accurately and rapidly find malignant tumor tissue, patient is timely controlled
Treat.Both can be applied in high-end accurate medical diagnosis, it is also possible to ultraviolet damage is predicted and detected in convenient, family
In be applied, be especially advantageous for popularization.
Brief description of the drawings
Fig. 1：Keratin 1 autofluorescence spectrogram in tissue.
Fig. 2：Action diagram of the Keratin 1 RNA interference for UV light-induced tissue autofluorescence.
Fig. 3：Keratin 1 RNA disturbs the action diagram for UV light-induced melanoma cells autofluorescence.
Specific embodiment
Using male C57 mouse, after treatment with ultraviolet light, mouse is raised in Animal House, and condition is 22-24 DEG C, 12
The light/dark circulation of hour, and can ad lib water intaking.After 6 hours, using laser confocal microscope to Keratin 1 in skin
Autofluorescence is imaged.Wherein the excitation wavelength of laser confocal microscope is 440-600 nm.
Using B16-F10 cells, cultivate at 37 degrees Celsius, in 5% carbon dioxide environment.The expression quantity of cytokeratin 1 is carried out
After interference is lowered 12 hours.Uv induction is carried out to cell that is interference and not interfering with.In 6 hours, laser co-focusing is used
Microscope imaging.Wherein the excitation wavelength of laser confocal microscope is 440-600 nm.
Fig. 1：The spectrogram of Keratin 1 in tissue, after UV light-induced, spatial distribution is not changed in.Fluorescence intensity strengthens.
Fig. 2：In tissue, it is found that ultraviolet light can induce the tissue autofluorescence of Keratin 1 expression high to strengthen.Disturbed using RNA
By the expression quantity reduction of Keratin 1, autofluorescence enhancing degree weakens method after uv induction.
Fig. 3：In melanoma cells model, the cellular autofluorescence enhancing that ultraviolet light can induce Keratin 1 to express is found.Utilize
By the expression quantity reduction of Keratin 1, autofluorescence enhancing degree weakens the method for RNA interference after uv induction.
Below will be by specifically describing, the present invention is further illustrated.
Unless otherwise defined, all technologies used herein and scientific terminology have general with the technical field of the invention
Logical technical staff is generally understood that identical implication.
Inventor has carried out substantial amounts of experiment, it is determined that tumor tissues keratin autofluorescence is significantly higher than normal structure, and this shows
As.
Experiment can show that tumor tissues keratin autofluorescence is significantly higher than normal structure more than.Therefore, Epidermal Keratin from
Fluorescence can be used as the mark of detection tumor tissues.
The present invention is based on this discovery, the method for establishing a kind of noninvasive markless detection tumor tissues of live body.The method includes
Following steps：
(1) after the tumor tissues in experimental subjects are subject to the light source comprising doses ultraviolet light to irradiate within 72 hours, detection
The experimental subjects passes through the autofluorescence of keratin in the tumor tissues that the light source containing ultraviolet light irradiates, wherein the dosage=
Uv power × irradiation time；
(2) detect wavelength that the experimental subjects sent by the tumor tissues that the light source containing ultraviolet light irradiates in 490-
Spontaneous fluorescence intensity in the range of 640nm；
(3) according to the method for the step (2), equally detect that the experimental subjects is subject to the swollen of the light source irradiation containing ultraviolet light
The autofluorescence intensity of knurl areas adjacent tissue；
(4) by the experimental subjects by the tumor tissues autofluorescence of the light source irradiation containing ultraviolet light and by containing ultraviolet light
The autofluorescence intensity of the tumor region tissue around of light source irradiation compares, and obtains strong through the autofluorescence of ultraviolet light induction
Degree rate of change；
(5) according to the keratin autofluorescence change rate of strength induced through ultraviolet light, it is right to be tested described in predicted detection
The tumor tissues position of elephant.
The mode of the utilization excitation tumor tissues keratin autofluorescence of aforementioned invention includes defeated using common continuous light
Go out excited mode, be modulated using electrical modulation in the mode that excites or the mode excited using pulse laser
At least one.
In foregoing detection method of the invention, preferably gland cancer and cutaneum carcinoma.
In foregoing detection method of the invention, it is preferred that lung, prostate, kidney, uterus, the tumor tissues on skin.
In foregoing detection method of the invention, a length of 200 nanometers -400 nanometers of ultraviolet light wave
In foregoing detection method of the invention, uv agent amount is 0-100 joules/square centimeter, preferably 0-20 joules/square
Centimetre, more preferably 0-10 joules/square centimeter.
In foregoing detection method of the invention, the wavelength preferably scope of 450-550 nm of the autofluorescence for being detected
It is interior, in the range of more preferably 460-500 nm.
Although those skilled in the art is it should be understood that for illustrative purposes, this document describes specific reality of the invention
Mode is applied, but various modifications can be carried out to it without departing from the spirit and scope of the present invention.Therefore, specific implementation of the invention
Mode and embodiment should not be considered as limiting the scope of the invention.The present invention is limited only by the appended claims.In the application
The all documents quoted are fully incorporated herein by reference.
Claims (10)
1. a kind of detection method of biomarker based on keratin autofluorescence as predicted detection tumour, including following step
Suddenly：
(1) after the tumor tissues in experimental subjects are subject to the light source comprising doses ultraviolet light to irradiate within 72 hours, detection
The experimental subjects passes through the autofluorescence of keratin in the tumor tissues that the light source containing ultraviolet light irradiates, wherein the dosage=
Uv power × irradiation time；
(2) detect wavelength that the experimental subjects sent by the tumor tissues that the light source containing ultraviolet light irradiates in 400-
Spontaneous fluorescence intensity in the range of 640nm；
(3) according to the method for the step (2), equally detect that the experimental subjects is subject to the swollen of the light source irradiation containing ultraviolet light
The autofluorescence intensity of knurl areas adjacent tissue；
(4) by the experimental subjects by the tumor tissues autofluorescence of the light source irradiation containing ultraviolet light and by containing ultraviolet light
The autofluorescence intensity of the tumor region tissue around of light source irradiation compares, and obtains strong through the autofluorescence of ultraviolet light induction
Degree rate of change；
(5) according to the keratin autofluorescence change rate of strength induced through ultraviolet light, it is right to be tested described in predicted detection
The malignant tumor tissue position of elephant.
2. the method for claim 1, it is characterised in that：The detection tumor tissues include biological tissue or frost sample
Product.
3. the method for claim 1, it is characterised in that：The tumor tissues include lung, liver, kidney, prostate, son
Palace, the tumour of skin part.
4. the method for claim 1, it is characterised in that：The type of the tumour is gland cancer and cutaneum carcinoma.
5. detected described in the method for Keratin 1 autofluorescence in sample, Keratin 1 is amplified by the ultraviolet light stimulus of doses
The signal of autofluorescence, uv agent amount is less than or equal between 100 joules/square centimeter.
6. the method for claim 1, it is characterised in that：Uv agent amount is less than or equal to 20 joules/square centimeter；It is preferred that
Less than or equal to 10 joules/square centimeter.
7. the method for claim 1, it is characterised in that：
The mode using excitation keratin autofluorescence includes the side excited using common continuous light output
Formula, at least one being modulated using electrical modulation in the mode that excites or the mode excited using pulse laser.
8. the method for claim 1, it is characterised in that：The keratin autofluorescence 1 is characterised by that spectral region exists
400-600nm。
9. method as claimed in claim 8, it is characterised in that：The Keratin 1 autofluorescence is characterised by that spectral region exists
450-550nm；Preferred spectral range is in 460-500nm.
10. it is a kind of based on Keratin 1 autofluorescence in tumor tissues after ultraviolet irradiation as detection Tumor biomarkers application.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109839507A (en) * | 2017-11-27 | 2019-06-04 | 江苏坤辉生物科技有限公司 | A kind of biomarker detecting ultraviolet light injury and its application |
| CN110068681A (en) * | 2018-01-24 | 2019-07-30 | 上海交通大学 | A kind of Novel marker predicted or detect lung cancer |
| CN110089999A (en) * | 2018-01-31 | 2019-08-06 | 上海交通大学 | Detection based on keratin protein fraction and keratin level and bilateral symmetry judges the application and method of cardiovascular and cerebrovascular disease |
| CN110151125A (en) * | 2018-02-12 | 2019-08-23 | 上海交通大学 | A kind of application and method based on nail spontaneous fluorescence intensity as biomarker in the product that preparation occurs for diagnosing |
| CN112710640A (en) * | 2019-10-25 | 2021-04-27 | 上海交通大学 | Method for evaluating interleukin level in serum based on epidermal keratin and fragment content |
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| CN101460850A (en) * | 2006-06-02 | 2009-06-17 | 阿特拉斯抗体有限公司 | Use of protein SATB2 as a marker for colorectal cancer |
| CN105852808A (en) * | 2015-05-08 | 2016-08-17 | 上海交通大学 | In-vivo non-invasive detection method for ultraviolet-light-induced skin injury and detection device adopted by same |
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| US5664574A (en) * | 1991-01-22 | 1997-09-09 | Non-Invasive Technology, Inc. | System for tissue examination using directional optical radiation |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109839507A (en) * | 2017-11-27 | 2019-06-04 | 江苏坤辉生物科技有限公司 | A kind of biomarker detecting ultraviolet light injury and its application |
| CN110068681A (en) * | 2018-01-24 | 2019-07-30 | 上海交通大学 | A kind of Novel marker predicted or detect lung cancer |
| CN110089999A (en) * | 2018-01-31 | 2019-08-06 | 上海交通大学 | Detection based on keratin protein fraction and keratin level and bilateral symmetry judges the application and method of cardiovascular and cerebrovascular disease |
| CN110151125A (en) * | 2018-02-12 | 2019-08-23 | 上海交通大学 | A kind of application and method based on nail spontaneous fluorescence intensity as biomarker in the product that preparation occurs for diagnosing |
| CN112710640A (en) * | 2019-10-25 | 2021-04-27 | 上海交通大学 | Method for evaluating interleukin level in serum based on epidermal keratin and fragment content |
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Application publication date: 20170613 |