JP3790792B2 - Simple determination method of risk of squamous cell carcinoma occurrence by exhalation and apparatus for the same - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The invention of this application relates to a simple determination method for determining a subject's risk of developing squamous cell carcinoma and a determination apparatus therefor. More specifically, the invention of this application relates to a simple determination method and a simple determination apparatus for determining a risk of developing squamous cell carcinoma of a subject from time-dependent changes in acetaldehyde concentration in the breath of the subject.
[0002]
[Prior art and its problems]
The number of deaths due to cancer was estimated at around 5.2 million worldwide in 1990, but it is estimated that it will approach 8 million in 2010 (Pisani, P., Parkin, DM, Bray, F., Ferlay, J., Int. J. Cancer 83, 18-29 (1999)), the number of cancer deaths in Japan approached 300,000 in 2000 (Ministry of Health, Labor and Welfare, 2000 vital statistics). This is more than the total number of deaths from cerebrovascular disease and deaths from heart disease. Moreover, by region, the number of trachea / bronchi and lung cancer is the largest, about 54,000, followed by gastric cancer with 50,000. Therefore, cancer countermeasures are positioned as one of the most important issues in disease countermeasures worldwide, and research on therapeutic drugs and treatment methods is being conducted from various angles.
[0003]
Recent research has led to a general recognition of the relationship between oral and pharyngeal cancer, esophageal cancer, lung cancer and smoking, skin cancer and ultraviolet light, etc. Statistics "'99). In recent years, the effects of drinking on various diseases have also been clarified. Regarding cancer, it has been clarified that excessive drinking increases the risk of oral and pharyngeal cancer and esophageal cancer. (E.g., HKSeitz, S. Matsuzaki, A. Yokoyama, N. Homann, S. Vakevainen, and XDWang, Alcoholism: Clinical and Experimental Research, Vol. 25, No. 5, 2001; S. Franceschi, F. Levi, C. LaVecchia, E. Conti, L. DalMaso, L. Barzan, and R. Talamini, Int. J. Cancer 83, 1-4, 1999). However, alcohol has long been referred to as the “Hundred Drugs” and the fact is that the relationship with carcinogenesis is generally not well recognized.
[0004]
Therefore, although the public's interest in cancer is high, there are currently no known specific preventive measures such as what kind of people should actually do what kind of health care. If there is a method for knowing whether an individual is in a risk group with a high risk of developing cancer, that is, a highly accurate method for determining and diagnosing a cancer risk group, doctors will provide guidance on preventive measures and the person's lifestyle habits. It is expected that awareness of improvement will increase.
[0005]
However, when such a diagnostic method involves complicated operations or pain of the subject, the cost is high, or if the diagnosis method has a long period of time, the penetration rate may be low.
[0006]
The invention of this application is to solve the above-mentioned problems and to provide a simple method for determining and diagnosing a high risk group of carcinogenesis due to alcohol, particularly a high risk group of squamous cell carcinoma occurrence. Yes.
[0007]
[Means for Solving the Problems]
In order to solve the problems as described above, the invention of this application firstly, after ingesting a certain amount of alcohol at a certain concentration, the time-dependent change in acetaldehyde concentration in exhaled breath was measured, and the acetaldehyde concentration rapidly increased. Provided is a simple method for determining the risk of developing squamous cell carcinoma, characterized by determining that the risk of developing squamous cell carcinoma is high when an increase is seen or the degree of decrease in acetaldehyde concentration over time is small .
[0008]
Secondly, according to the invention of this application, after ingesting a certain amount of alcohol at a certain concentration, the time-dependent change in acetaldehyde concentration in the breath is measured, and when the acetaldehyde concentration does not become 10 nmol or less within 30 minutes, squamous cell carcinoma Provided is a simple determination method for the risk of occurrence of squamous cell carcinoma determined to have a high risk of occurrence.
[0009]
Third, the invention of this application is a simple determination device for determining the risk of occurrence of squamous cell carcinoma by exhalation, and at least,
A breath induction part that introduces exhaled breath from a subject after ingesting a constant amount of alcohol, an acetaldehyde concentration measurement part that measures the acetaldehyde concentration of the introduced breath over time, and squamous epithelium from changes in acetaldehyde concentration over time A risk level determination unit for determining the risk level of cancer occurrence and an index for this determination, a risk level index setting unit to be transmitted to the risk level determination unit,
A determination display unit that displays a determination result from the risk determination unit,
In the risk index setting unit, when the acetaldehyde concentration shows a rapid increase in the course of change over time or a decrease level smaller than a predetermined decrease level, the risk level is set as a high risk level. An apparatus for simple determination of the risk of occurrence of epithelial cancer is provided.
[0010]
The fourth aspect of the present invention also provides the above-described apparatus for simple determination of the risk of occurrence of squamous cell carcinoma, wherein the acetaldehyde concentration is reduced to 10 nmol or less within 30 minutes.
[0011]
DETAILED DESCRIPTION OF THE INVENTION
The inventors of this application have so far studied the mechanism of carcinogenesis, particularly in multiple cancer patients with both head and neck cancer and esophageal cancer. He also discovered and reported that multiple atypical epithelium was observed in the esophageal mucosa of patients with both head and neck cancer and esophageal cancer (Muto et al. Gut. 2000 Aug; 47 (2 ): 256-261).
[0012]
According to the inventors' earnest studies, such multiple atypical epithelium is seen only in patients who prefer alcohol among multiple cancer patients, suggesting a relationship between multiple atypical epithelial cancer and alcohol drinking. However, about 2/3 patients who did not have multiple atypical epithelium at all even if they consumed a large amount of alcohol, and there was no relationship between alcohol intake and the occurrence of multiple atypical epithelium.
[0013]
Accordingly, the inventors of this application focused on the patient's ability to metabolize alcohol, and examined gene polymorphisms of two enzymes involved in alcohol metabolism, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Each enzyme is known to vary depending on the gene polymorphism, but the inventors have found that the frequency of occurrence of multiple atypical epithelium varies greatly depending on the combination of the gene polymorphisms of these two enzymes.
[0014]
It is known that human alcohol dehydrogenase (ADH) has three isomers, ADH1, ADH2 and ADH3. Among them, ADH3 includes ADH3-1 allyl homozygote (ADH3 * 1/3 * 1) having a high alcohol metabolism rate and ADH3-2 allyl (ADH3 * 1/3 * 2 and ADH3 * 2 /) having a low alcohol metabolism rate. 3 * 2) exists. (AFOlshan, MCWeissler, MAWatson and DABell; Carcinogenesis vol.22 no.1 pp.57-61, 2001)
On the other hand, aldehyde dehydrogenase (ALDH), which further decomposes acetaldehyde produced by alcohol decomposition, is known to have ALDH1 that works when aldehyde is at a high concentration and ALDH2 that can also decompose a low concentration of aldehyde. Yes.
[0015]
ALDH2 gene polymorphism includes active ALDH2-1 allele (ALDH2 * 1/2 * 1) and inactive ALDH2-2 allele (ALDH2 * 1/2 * 2 and ALDH2 * 2/2 * 2), It is known that a person who is prone to a flushing reaction such as redness of the face due to drinking has ALDH2-2 allyl. Moreover, ALDH2-2 allyl is not found in Westerners and is known to exist only in Asian ethnic groups. (HKSeitz, S. Matsuzaki, A. Yokoyama, N. Homann, S. Vakevainen, and XDWang; Alcoholism: Clinical and Experimental Research, Vol. 25, No. 5)
The inventors of this application have found that about 85% of patients with both ADH3-2 allyl and ALDH2-2 allele have multiple atypical epithelium in patients with double cancer, and ADH3-2 allele in the cell chromosome and The present invention provides a method for determining a group of multiple squamous cell carcinoma risks that detects ALDH2-2 allele.
[0016]
Such a determination method is very accurate, and it can be easily accepted by society if it is implemented as an item for cancer screening in the current “clinical checkup” and “regular health checkup”. It is very useful because it is thought to improve general awareness.
[0017]
However, for actual diagnosis, for example, normal cells are collected from the throat and esophagus by endoscopy, DNA is extracted from peripheral blood lymphocytes, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), etc. There are multiple steps such as detection of ADH3-2 allyl and ALDH2-2 allyl, and several days are required for the determination. Further, since an analyzer for gene diagnosis is required, the diagnosis cost is likely to be high, and it is difficult to make a quick determination at a small clinic or clinic. In addition, many companies, etc., conduct health checkups, including physical examinations and endoscopy, only for employees over a certain age. Not very much. However, young cancer patients are also increasing, and cancer prevention and lifestyle improvement are important issues regardless of age. Therefore, it would be still preferable if there is a method for screening a squamous cell cancer risk group more easily without performing a genetic test.
[0018]
The inventors of the present invention have found that there is a correlation between the combination of ADH3 and ALDH2 gene polymorphisms when the acetaldehyde concentration in exhaled breath was measured by alcohol loading a patient who had both head and neck cancer and esophageal cancer. The present invention has been achieved.
[0019]
That is, in the simple determination method of the squamous cell carcinoma occurrence risk of the invention of this application, after ingesting a certain amount of alcohol at a certain concentration, the time-dependent change in acetaldehyde concentration in the breath is measured, and whether the aldehyde concentration rapidly increases When the degree of decrease over time is small, it is determined that the risk of occurrence of squamous cell carcinoma is high.
[0020]
In such a simple determination method, alcohol previously ingested by the subject is decomposed in the body and converted into acetaldehyde and discharged into the breath. At that time, when a rapid increase in acetaldehyde concentration immediately after alcohol intake is observed, or when the degree of decrease in acetaldehyde concentration with time is small, it can be determined that both ADH3-2 allyl and ALDH2-2 allyl are present. .
[0021]
According to the study by the inventors, the concentration of acetaldehyde in exhaled breath when ingesting an equal amount of alcohol over the same period of time is determined by the combination of genetic polymorphisms in the subject: ADH3-2 (−) / ALDH2-2 (− ), ADH3-2 (+) / ALDH2-2 (−), ADH3-2 (−) / ALDH2-2 (+), ADH3-2 (+) / ALDH2-2 (+) in this order. As is clear from the examples described later, the difference is clear.
[0022]
Therefore, a reference value is provided, and it may be determined that the risk of developing squamous cell carcinoma is high when the aldehyde concentration in exhaled air is equal to or higher than the reference value, or when it does not become lower than the reference value within a certain time. The reference value for such determination is that the combination of genetic polymorphisms is ADH3-2 (−) / ALDH2-2 (−), ADH3-2 (+) / ALDH2-2 (−), ADH3-2 (−). It is preferable that the subjects who are / ALDH2-2 (+) and ADH3-2 (+) / ALDH2-2 (+) are comprehensively measured under the same conditions and set based on the results. For example, based on the research results of the inventors, if the acetaldehyde concentration in exhaled breath does not become 10 nmol or less after 30 minutes have elapsed after ingesting 200 ml of 6% alcohol, it is determined that the risk of squamous cell carcinoma is high it can. Of course, the reference value may be set as appropriate according to the age, sex, race, etc. of the subject.
[0023]
The invention of this application further provides a simple determination device for determining the risk of squamous cell carcinoma from the concentration of acetaldehyde in exhaled breath. Such a simple determination device has a configuration as shown in FIG. That is, at least
(A) an exhalation introduction unit for introducing exhalation from a subject after ingesting a certain amount of alcohol at a certain concentration, (b) an acetaldehyde concentration measurement unit for measuring the acetaldehyde concentration of the introduced exhalation over time, and (c) A risk determination unit for determining the risk of squamous cell carcinoma from a change in acetaldehyde concentration with time, and (d) a risk index setting unit for setting an index for this determination and communicating it to the risk determination unit; ,
(E) It shall have a determination display part which displays the determination result from a risk determination part. In such a simple determination device, in the risk index setting unit (d), when the acetaldehyde concentration rapidly increases in the course of change with time, or when the degree of decrease in acetaldehyde concentration is smaller than a predetermined value, It is set as an index of “high risk”.
[0024]
In the apparatus for simple determination of the risk of occurrence of squamous cell carcinoma according to the invention of this application, exhalation is performed by injecting a predetermined amount of alcohol into a subject and then breathing into a commercially available sample bag for gas sampling, for example. Can be collected. Alternatively, in the case where the breath introduction unit of the simple determination device has a mouthpiece that can directly breathe, the subject may breathe in by placing the mouth on the mouthpiece. At this time, the amount of exhaled air collected is not particularly limited. The amount may be an amount that does not place a large burden on the subject, and can be appropriately changed according to the detection sensitivity of the acetaldehyde concentration measurement unit.
[0025]
In the case of collecting exhaled gas into the sample bag, the exhalation introducing part (a) in the apparatus for simple determination of the invention of this application can extract exhaled gas from the sample bag with a syringe, insert the syringe, and inject the collected exhaled gas It may be an insertion inlet, or may have a structure that allows the sample bag to be directly connected via a stopcock or the like. On the other hand, when inhaling exhaled breath directly into the device for simple determination, the mouthpiece may be a small diameter tube or straw that is contained in the mouth and breathes in, or a face mask that breathes into the mouth. There may be. It is preferable that these can be removed from the viewpoint of hygiene, can be sterilized, or disposable. Of course, these sample bags and mouthpieces may have filters, stopcocks, one-way valves, and the like.
[0026]
The introduced exhaled air is then transferred to the acetaldehyde concentration measurement unit (b), and the acetaldehyde concentration is measured over time. At this time, exhaled air may be transferred from the exhalation introduction unit to the acetaldehyde concentration measurement unit using an inert gas such as nitrogen or argon as a carrier gas, or exhalation may be convection and diffusion as it is. Such an acetaldehyde concentration measuring unit may have any configuration and structure as long as it can measure the concentration of acetaldehyde, and is not particularly limited. Specifically, a Breath Gas Analyzer improved for acetaldehyde, gas chromatography, a gas detector tube, a reagent-coated crystal tensioning device, and the like are exemplified.
[0027]
Breath Gas Analyzer is a device that measures the concentration of hydrogen and methane in exhaled breath, which has been used for diagnosis of lactose digestion and absorption disorders and diabetic gastrointestinal disorders. If this detection part is improved for acetaldehyde, it can be applied as an acetaldehyde concentration measurement part (b) for the simple determination device of the present invention. On the other hand, gas chromatography is based on the difference in the absorption and desorption speed of each component when a column made of stainless steel or glass is filled with a filler such as molecular sieves, silica gel, or porous resin beads. This is a method for separating, analyzing, and quantifying gases, and various types of column packs suitable for analyzing acetaldehyde can be selected. The gas detector tube is a simple method of analyzing the concentration based on the color tone change caused by the reaction between a specific reagent and a gas, and the measurement range of the acetaldehyde concentration of 0.1 ppm to 20 ppm by the color tone change such as yellow → red. A wide variety of products such as those that can be measured for 1 to 10 hours are commercially available. Since the gas detection tube cannot be used repeatedly, it is disposable every time, but it can be said to be a simple and effective means if an amount of exhalation corresponding to the measurement range can be collected. Furthermore, the reagent-applied quartz crystal slidator applies a reagent such as a Schiff base that specifically reacts with acetaldehyde to the surface of the vibrator and detects a change in oscillation frequency that occurs when acetaldehyde reacts. In the invention of this application, gas chromatography having high measurement accuracy and a wide measurement concentration range is preferably used as the acetaldehyde concentration measurement unit.
[0028]
The measurement result in the acetaldehyde concentration measurement unit (b) as described above is transmitted to the risk determination unit (c) as an electric signal or the like by the signal transmission means. Then, in this risk determination unit (c), the risk of occurrence of squamous cell carcinoma is determined from the change in the acetaldehyde concentration with time.
[0029]
Such a risk determination unit (c) can be, for example, a computer, and the value of the acetaldehyde concentration in the breath measured by the acetaldehyde concentration measurement unit (b) over time is set as a risk index setting part (d). It is possible to compare and judge the index set as input. The risk index input and set in the risk index setting part (d) is a combination of genetic polymorphisms of ADH3-2 (−) / ALDH2-2 (−), ADH3-2 (+) / ALDH2-2. (−), ADH3-2 (−) / ALDH2-2 (+), ADH3-2 (+) / ALDH2-2 (+) subjects were measured comprehensively under the same conditions, and the results were Can be set to the original. In addition, the setting can be changed at any time according to the age, sex, race, etc. of the subject, and is not particularly limited.
[0030]
In such a risk index setting part (d), when the acetaldehyde concentration shows a rapid increase in the process of change with time or a decrease degree smaller than a predetermined decrease degree, the risk degree is set to be high. According to the earnest study by the inventors, it can be set that the risk of squamous cell carcinoma is high when the concentration of acetaldehyde in the breath does not become 10 nmol or less after 30 minutes have elapsed after ingesting 200 ml of 6% alcohol.
[0031]
The determination result is further transmitted to the determination display section (e) by the signal transmission means and displayed. At this time, as the judgment display part (e), a numerical value or graph is displayed as an image like a liquid crystal monitor or a CRT monitor, a sound is output like a speaker, and a printer is used. Examples where the result is printed on the paper are illustrated.
[0032]
The apparatus for simple determination of the risk of occurrence of squamous cell carcinoma as described above has at least the configuration as described above, but also emits voice, characters, signals, images, etc. instructing the subject to breathe in. You may have a display means etc. which show a test means and a measurement progress to a test subject or an operator. Such a simple determination device can be moved and carried by reducing the size of each component. This makes it possible to measure the risk of epithelial cancer occurrence quantitatively, quickly, simply, and with high accuracy.
[0033]
【Example】
<Example 1>
78 subjects who passed 2 hours or more after meals were ingested with 200 ml of grapefruit juice containing 6% alcohol over 10 minutes. Of the 78, 26 were unable to participate in the test due to strong alcohol intolerance.
[0034]
Therefore, for the remaining 52 people, exhaled breath was collected with an exhalation collection pack before alcohol intake, immediately after ingestion, 10 minutes later and 30 minutes later, and gas chromatography (column: Polapack Q60 / 80 glass column 3 mm × 2 m; GL Science Acetaldehyde concentration in 1 ml of exhaled breath was analyzed by a company, detector: FID, carrier gas: helium, flow rate: 40 mlmin.
[0035]
The results are shown in FIG.
[0036]
Acetaldehyde, which was not detected at all before alcohol intake, was detected immediately after drinking. Moreover, the difference was seen by the peak value and the value after 10 minutes and 30 minutes by the combination of gene polymorphism allele.
[0037]
Specifically, the combinations of gene polymorphisms are ADH3-2 (−) / ALDH2-2 (−), ADH3-2 (+) / ALDH2-2 (−), ADH3-2 (−) / ALDH2-2 ( +) And ADH3-2 (+) / ALDH2-2 (+) in the order of increasing acetaldehyde concentration.
[0038]
Since the risk of occurrence of multiple atypical epithelium also increased significantly in the same order, it was shown that the risk of occurrence of squamous cell carcinoma can be accurately predicted by measuring the concentration of acetaldehyde in exhaled breath. It was also shown that high risk groups with multiple cancers can be found with high accuracy. That is, as shown in FIG. 2, when the combinations of gene polymorphisms are ADH3-2 (−) / ALDH2-2 (−) and ADH3-2 (+) / ALDH2-2 (−), 30 minutes later It was determined that the acetaldehyde concentration in exhaled breath was 10 nmol or less and the risk of squamous cell carcinoma was low. On the other hand, when the combination of gene polymorphisms is ADH3-2 (−) / ALDH2-2 (+) and ADH3-2 (+) / ALDH2-2 (+), the concentration of acetaldehyde in exhaled breath is 10 nmol even after 30 minutes. Since it was not below (both were maintained at 50 nmol or more), it could be determined that the risk of squamous cell carcinoma was high.
[0039]
【The invention's effect】
As described in detail above, the invention of this application provides a simple determination method and determination apparatus that allow a subject to diagnose the risk of developing squamous cell carcinoma by measuring the aldehyde concentration in exhaled breath.
[0040]
Such a method for determining the risk of squamous cell carcinoma development does not require genetic testing, enables quantitative and non-invasive evaluation in a short time, and requires expensive genetic analysis equipment and complicated operations Therefore, it can be easily introduced in small clinics and clinics, is easily accepted by society, and is expected to raise social interest in cancer prevention.
[0041]
In addition, the device for determining the risk of occurrence of squamous cell carcinoma of the present application not only makes it possible to easily measure the risk of the occurrence of squamous cell carcinoma, but also makes it possible to obtain a portable device for simple determination by downsizing. . With such a small simple determination device, it becomes possible to quickly determine the risk of squamous cell carcinoma occurrence at any medical site, and it is expected that cancer prevention will more commonly permeate.
[0042]
If the subject himself / herself recognizes whether or not the risk of carcinogenesis is high, it becomes possible to tackle specific and effective cancer prevention, and thus the present invention is highly useful.
[Brief description of the drawings]
FIG. 1 is a schematic diagram showing an outline of a device for simple determination of the risk of occurrence of squamous cell carcinoma in the invention of this application.
FIG. 2 is a graph showing the relationship between the acetaldehyde concentration in exhaled breath due to alcohol intake and the combination of ADH3 and ALDH2 gene polymorphisms in the examples of the invention of this application.

Claims (4)

After ingesting a certain amount of alcohol at a certain concentration, measure changes over time in the acetaldehyde concentration in exhaled breath. A simple determination method for the risk of occurrence of squamous cell carcinoma, characterized by determining that the risk of occurrence of epithelial cancer is high. Claims: After ingesting a certain amount of alcohol at a certain concentration, the time-dependent change in acetaldehyde concentration in exhaled breath is measured, and if the acetaldehyde concentration does not fall below 10 nmol within 30 minutes, it is determined that the risk of developing squamous cell carcinoma is high 1. A simple determination method for the risk of squamous cell carcinoma occurrence of 1. A simple determination device for determining the risk of squamous cell carcinoma occurrence by exhalation, at least,
A breath induction part that introduces exhaled breath from a subject after ingesting a constant amount of alcohol, an acetaldehyde concentration measurement part that measures the acetaldehyde concentration of the introduced breath over time, and squamous epithelium from changes in acetaldehyde concentration over time A risk level determination unit for determining the risk level of cancer occurrence and an index for this determination, a risk level index setting unit to be transmitted to the risk level determination unit,
A determination display unit that displays a determination result from the risk determination unit,
In the risk index setting unit, when the acetaldehyde concentration shows a rapid increase in the course of change over time or a decrease level smaller than a predetermined decrease level, the risk level is set as a high risk level. Device for simple determination of epithelial cancer risk. The apparatus for simple determination of the risk of occurrence of squamous cell carcinoma according to claim 3, wherein the predetermined decrease is that the acetaldehyde concentration becomes 10 nmol or less within 30 minutes.

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