JPH06103303B2 - How to distinguish malignant tumor - Google Patents
How to distinguish malignant tumorInfo
- Publication number
- JPH06103303B2 JPH06103303B2 JP1278623A JP27862389A JPH06103303B2 JP H06103303 B2 JPH06103303 B2 JP H06103303B2 JP 1278623 A JP1278623 A JP 1278623A JP 27862389 A JP27862389 A JP 27862389A JP H06103303 B2 JPH06103303 B2 JP H06103303B2
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- JP
- Japan
- Prior art keywords
- sta
- tissue
- lectin
- malignant tumor
- tissues
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は、腫瘍細胞が良性か悪性か否かを判別する悪性
腫瘍の判別方法に関するものである。TECHNICAL FIELD The present invention relates to a method for discriminating malignant tumors, which discriminates whether tumor cells are benign or malignant.
（発明の背景） 細胞質や細胞膜の糖鎖構造が癌化に伴なって変化するこ
とが知られている。このような糖鎖構造の変化に基づ
き、特定のオリゴ糖と強く反応するレクチンがしばしば
用いられている。しかし従来知られているレクチンは何
れも癌の種類に特異的なものであり、癌組織であるか否
かを判別するのに広く一般的に使用できるものはなかっ
た。(Background of the Invention) It is known that the sugar chain structure of the cytoplasm and the cell membrane changes with canceration. Lectins that strongly react with specific oligosaccharides are often used based on such changes in sugar chain structure. However, all of the conventionally known lectins are specific to the type of cancer, and none of them can be widely and generally used to determine whether or not they are cancer tissues.
癌細胞であるか否かは、組織標本を形態学的に観察すれ
ば、正常組織の組織像との相違から或る程度は判定出来
る。しかしながら、良性腫瘍の中には、悪性腫瘍と見分
けにくいものもあり、単なる形態学的な観察だけでは十
分ではなかった。Whether or not it is a cancer cell can be determined to some extent by morphologically observing a tissue specimen from the difference from the tissue image of a normal tissue. However, some benign tumors are difficult to distinguish from malignant tumors, and mere morphological observation was not sufficient.
このような状況下において、本発明者は、種々のレクチ
ンについて癌組織との反応性を調べたところ、STA（ジ
ャガイモレクチン:Solanum Tuberosum Lectin）が上皮
性、間葉性を問わず多くの癌細胞と結合すること、正常
細胞や良性腫瘍のものとはほとんど結合しないことを見
出した。本発明はこのような知見に基づいて完成された
ものである。Under such circumstances, the present inventor examined the reactivity of various lectins with cancer tissue, and found that STA (potato lectin: Solanum Tuberosum Lectin) showed many cancer cells regardless of epithelial or mesenchymal. It was found that it binds to and that of normal cells and benign tumors. The present invention has been completed based on such findings.
（発明の目的） すなわち本発明は、腫瘍組織が良性か悪性か否かを判別
する悪性腫瘍の判別方法を提供することを目的とする。(Object of the Invention) That is, an object of the present invention is to provide a method for determining a malignant tumor, which determines whether the tumor tissue is benign or malignant.
（発明の構成） このような本発明の目的は、腫瘍組織標品をジャガイモ
レクチン（Solanum Tuberosum Lectin）で処理し、ジャ
ガイモレクチンの結合の有無を形態学的に観察すること
を特徴とする悪性腫瘍の判別方法により達成される。(Structure of the Invention) An object of the present invention is to treat a tumor tissue preparation with potato lectin (Solanum Tuberosum Lectin), and morphologically observe the presence or absence of the binding of the potato lectin to a malignant tumor. It is achieved by the discrimination method of.
STA（ジャガイモレクチン:Solanum Tuberosum Lectin）
はN-アセチル−Ｄ−グルコサミンに特異的に結合するレ
クチンとして知られている。この糖鎖が悪性腫瘍でのみ
検出できる理由は明らかではないが、STAが主として小
胞体やゴルジ体など細胞内物質と結合することから、細
胞膜表面よりもむしろ細胞内に多く存在すると思われ
る。またN-アセチル−Ｄ−グルコサミンは、細胞膜表面
に多く分布することが知られているＮ−アセチルノイラ
ミン酸やムコポリサッカライド（ヒアルロン酸やコンド
ロイチン等）の生合成過程における前駆体であるから、
癌化に伴なう細胞膜表面の糖鎖構造の変化と、その糖鎖
構成分子の細胞内合成の変動を反映していると思われ
る。STA (potato lectin: Solanum Tuberosum Lectin)
Is known as a lectin that specifically binds to N-acetyl-D-glucosamine. The reason why this sugar chain can be detected only in malignant tumors is not clear, but STA mainly binds to intracellular substances such as the endoplasmic reticulum and the Golgi apparatus, and therefore, it is considered that it is more abundant in the cells than on the cell membrane surface. In addition, N-acetyl-D-glucosamine is a precursor in the biosynthesis process of N-acetylneuraminic acid and mucopolysaccharide (hyaluronic acid, chondroitin, etc.), which are known to be widely distributed on the cell membrane surface,
It seems to reflect changes in sugar chain structure on the cell membrane surface associated with canceration and changes in intracellular synthesis of sugar chain constituent molecules.
本発明の使用するSTAは市販のものを用いることができ
例えば宝酒造（株）やEYラボラトリー社のものを用いる
ことができる。As the STA used in the present invention, a commercially available STA can be used, and for example, those from Takara Shuzo Co., Ltd. or EY Laboratory Co. can be used.
本発明は、常法により作製した組織標品切片または細胞
懸濁液を、常法（Biochem.J.,135,307（1973））に従い
STAで処理し、鏡検によりSTA結合の有無、その分布を観
察することにより、悪性腫瘍であるか否かを判定する。
STA処理は、通常、組織標品切片または細胞懸濁液に最
終濃度25μg/mlのSTAを添加し、室温で30〜60分間反応
させることにより行なう。The present invention uses a tissue preparation section or cell suspension prepared by a conventional method according to a conventional method (Biochem. J., 135 , 307 (1973)).
By treating with STA and observing the presence or absence of STA binding and its distribution by microscopic examination, it is determined whether or not it is a malignant tumor.
The STA treatment is usually carried out by adding STA at a final concentration of 25 μg / ml to a tissue preparation section or a cell suspension and allowing the reaction at room temperature for 30 to 60 minutes.
この際、ローダミンやテキサスレッド等の色素やFITCな
どの蛍光色素で標識したSTAを用いれば、形態学的観察
は容易となる。あるいはペルオキシダーゼやアルカリフ
ォスファターゼなどの酵素やビオチンなど標識したSTA
を用い、STA処理後にその酵素反応を利用した特殊染色
を行なってもよい。この際には観察を容易にするため核
染色を行なってもよい。またこのような標識STAを使わ
ない場合には、STA処理後に抗STA抗体で処理し、ペルオ
キシダーゼ・抗ペルオキシダーゼ法などにより特殊染色
する免疫組織化学的観察を行なってもよい。At this time, if STA labeled with a dye such as rhodamine or Texas red or a fluorescent dye such as FITC is used, morphological observation becomes easy. Alternatively, an enzyme such as peroxidase or alkaline phosphatase, or STA labeled with biotin, etc.
It is also possible to perform special staining using the enzyme reaction after the STA treatment by using. At this time, nuclear staining may be performed to facilitate observation. When such a labeled STA is not used, it may be treated with an anti-STA antibody after the STA treatment and subjected to immunohistochemical observation with special staining by a peroxidase / antiperoxidase method or the like.
電子顕微鏡によりSTA結合の有無を観察する場合には、
フェリチン標識したSTAを用いるのが好ましい。この場
合には通常の電子顕微鏡標品の作製手順に従って鏡検試
料を作製する。When observing the presence or absence of STA binding with an electron microscope,
It is preferable to use ferritin-labeled STA. In this case, a microscopic sample is prepared according to the usual procedure for preparing an electron microscope standard.
（実施例） ヒト甲状腺の乳頭腺癌（悪性腫瘍）と濾胞腺腫（良性腫
瘍）についてSTA処理を行なった。(Example) STA treatment was performed on papillary adenocarcinoma (malignant tumor) and follicular adenoma (benign tumor) of human thyroid.
外科手術から得た各組織を10％緩衝ホルマリン溶液で固
定しパラフィン包埋してミクロトーム切片（厚さ４μ
ｍ）を調製した。脱パラフィン後PBSで数回洗浄し、0.3
％H2O2−メタノール溶液に20分間浸漬して組織に内在す
るペルオキシダーゼを不活化した。この標品を１％BSA
（牛血清アルブミン）溶液で５分間処理し、HRP−STA
（西洋ワサビペルオキシダーゼ標識ジャガイモレクチ
ン:EY−ラボラトリー社製）溶液の40倍希釈液に浸漬し
室温で30分間処理した。次いでこの組織切片を１％BSA
溶液で５分間処理後PBSで洗浄し、0.01％3,3′−ジアミ
ノベンジジン−50mMTris-HCl緩衝液（pH7.6）に３〜10
分間浸して発色させた。発色後、ヘマトキシリンにより
核染色した。この後蒸留水で洗浄、さらに脱水して、ス
ライドガラスに封入し、25倍で鏡検した。Each tissue obtained from surgery was fixed with 10% buffered formalin solution, embedded in paraffin, and microtome section (thickness 4 μm).
m) was prepared. After deparaffinization, wash with PBS several times to 0.3
The peroxidase existing in the tissue was inactivated by immersing it in a% H 2 O 2 -methanol solution for 20 minutes. This standard is 1% BSA
Treatment with (bovine serum albumin) solution for 5 minutes, HRP-STA
(Horse horseradish peroxidase-labeled potato lectin: manufactured by EY-Laboratory) It was immersed in a 40-fold diluted solution and treated at room temperature for 30 minutes. Then, this tissue section is cut with 1% BSA.
After treatment with the solution for 5 minutes, the plate was washed with PBS, and added with 0.01% 3,3'-diaminobenzidine-50 mM Tris-HCl buffer (pH 7.6) at 3 to 10%.
Soak for a minute to develop color. After color development, nuclear staining was performed with hematoxylin. After that, it was washed with distilled water, further dehydrated, enclosed in a slide glass, and examined microscopically at 25 times.
第１図に示すように、甲状腺の乳頭腺癌（同図中央部）
では乳頭状の細胞増生からなる結節が見られ、その細胞
質は褐色に染色され、びまん性にSTA陽性であった。同
図周辺部に存在する正常組織ではSTA陰性であった。As shown in Fig. 1, papillary adenocarcinoma of the thyroid gland (center part of the same figure)
A papillary nodule consisting of hyperplasia was observed in the cytoplasm, and its cytoplasm was stained brown and was diffusely STA-positive. STA was negative in the normal tissues around the same figure.
一方、良性腫瘍である濾胞腺腫では染色部位が見当たら
ずSTA陰性であった（第２図）。On the other hand, in the benign tumor, follicular adenoma, no staining site was found and it was STA-negative (Fig. 2).
このようにSTAは悪性腫瘍に陽性で、良性腫瘍には陰性
であった。Thus, STA was positive for malignant tumors and negative for benign tumors.
第３図は、同様の実験をヒトの胃の高分化腺癌組織につ
いて行なった場合の細胞組織像（50倍）である。図中矢
印で示した左半分には癌組織が観察され、その細胞質は
びまん性にSTA陽性であった。図中右半分は正常の幽門
腺であり、STA陰性であった。FIG. 3 is a cell tissue image (50 times) when the same experiment was performed on a well-differentiated adenocarcinoma tissue of human stomach. Cancer tissue was observed in the left half indicated by the arrow in the figure, and its cytoplasm was diffusely STA-positive. The right half of the figure was a normal pyloric gland, which was STA negative.
第４図は、同じくヒト大腸の高分化腺癌組織の細胞組織
像（25倍）であり、図面左下部分（図面矢印部分）の癌
組織はSTA陽性であり、図面上部1/4に見られる正常粘膜
組織ではSTA陰性であった。Fig. 4 is also a cell histology of a well-differentiated adenocarcinoma tissue of human colon (25 times), the cancer tissue in the lower left portion of the drawing (arrow portion in the drawing) is STA-positive, and is seen in the upper 1/4 of the drawing. STA was negative in normal mucosal tissues.
同様なSTA染色を上皮性、間葉性等の種々の悪性腫瘍組
織約110例について行なったところ、何れの悪性腫瘍組
織もSTA陽性であり、STAは悪性腫瘍組織に広く特異的に
結合すること、すなわち偽陰性は殆ど存在しないことが
わかった。調べた悪性腫瘍組織の結果の一部を下記の表
に示す。Similar STA staining was performed on about 110 cases of various malignant tumor tissues such as epithelial and mesenchymal tissues. All malignant tumor tissues were STA positive, and STA was widely and specifically bound to malignant tumor tissues. That is, it was found that there are almost no false negatives. Some of the results of the malignant tumor tissues examined are shown in the table below.
また各種正常組織についてSTA染色したところ、胃粘
膜、大腸粘膜上層部、前立腺、乳腺、肝細胞などではST
A陽性となったが、肝細胞以外では極めて軽微にしか染
色されず、殆んどの臓器組織では正常細胞と癌組織とは
STA染色により判別できることがわかった。 In addition, STA staining of various normal tissues revealed that ST was found in the gastric mucosa, colon mucosa upper layer, prostate, mammary gland, hepatocytes, etc.
Although it became A-positive, it was stained only very slightly except in hepatocytes, and normal cells and cancer tissues were not found in most organ tissues.
It was found that it could be identified by STA staining.
さらに病理学的に良性腫瘍と診断された臓器組織約30例
について調べたところSTA陽性となるものは皆無であっ
た。Furthermore, when we examined about 30 organ tissues pathologically diagnosed as benign tumors, none of them became STA-positive.
このようにSTAを用いた本発明方法は、癌組織をほぼ確
実に認識できる。Thus, the method of the present invention using STA can almost certainly recognize a cancer tissue.
（発明の効果） 以上のように本発明は、腫瘍組織標品をSTA（ジャガイ
モレクチン）で処理し、STAの結合の有無を形態学的に
観察するものであり、悪性腫瘍と良性腫瘍とを容易に判
別することができる。(Effect of the invention) As described above, the present invention is to treat a tumor tissue preparation with STA (potato lectin) and morphologically observe the presence or absence of STA binding. It can be easily identified.
第１図は本発明の方法により処理したヒト甲状腺の乳頭
腺癌の組織像を示す図、第２図は同じくヒト甲状腺の濾
胞腺腫（良性腫瘍）の組織像を示す図、第３図は同じく
ヒト胃の高分化腺癌の組織像を示す図、第４図は同じく
ヒト大腸の高分化腺癌の組織像を示す図である。FIG. 1 is a diagram showing a histological image of papillary adenocarcinoma of human thyroid treated by the method of the present invention, FIG. 2 is a diagram showing a histological image of follicular adenoma (benign tumor) of human thyroid gland, and FIG. 3 is also the same. FIG. 4 is a diagram showing a histological image of well-differentiated adenocarcinoma of human stomach, and FIG. 4 is a diagram showing a histological image of well-differentiated adenocarcinoma of human colon.
Claims (2)
num Tuberosum Lectin）で処理し、ジャガイモレクチン
の結合の有無を形態学的に観察することを特徴とする悪
性腫瘍の判別方法。1. A tumor tissue preparation is prepared from potato lectin (Sola).
num tuberosum lectin), and morphologically observing the presence or absence of potato lectin binding.
おり、ジャガイモレクチンの結合の有無は酵素組織化学
的に観察することを特徴とする請求項（１）記載の悪性
腫瘍の判別方法。2. The method for discriminating malignant tumors according to claim 1, wherein the potato lectin is enzyme-labeled in advance, and the presence or absence of binding of the potato lectin is observed by enzyme histochemistry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1278623A JPH06103303B2 (en) | 1989-10-27 | 1989-10-27 | How to distinguish malignant tumor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1278623A JPH06103303B2 (en) | 1989-10-27 | 1989-10-27 | How to distinguish malignant tumor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH03140865A JPH03140865A (en) | 1991-06-14 |
| JPH06103303B2 true JPH06103303B2 (en) | 1994-12-14 |
Family
ID=17599861
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1278623A Expired - Lifetime JPH06103303B2 (en) | 1989-10-27 | 1989-10-27 | How to distinguish malignant tumor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH06103303B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002077649A1 (en) * | 2001-03-27 | 2002-10-03 | Teikoku Hormone Mfg. Co., Ltd. | Method of diagnosing breast cancer |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5729951A (en) * | 1980-07-30 | 1982-02-18 | Nippon Koutai Kenkyusho:Kk | Determintion of sugar side chain related to cancer |
| JPS62257063A (en) * | 1986-05-01 | 1987-11-09 | Kokichi Sugano | Method for measuring cancer related glycoprotein |
| JPS63152998A (en) * | 1986-12-16 | 1988-06-25 | Konica Corp | Determination of sugar transferase |
-
1989
- 1989-10-27 JP JP1278623A patent/JPH06103303B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH03140865A (en) | 1991-06-14 |
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