實(shí)驗(yàn)性肝癌癌組織的發(fā)生和甲胎蛋白表達(dá)

1、    實(shí)驗(yàn)性肝癌癌組織的發(fā)生和甲胎蛋白表達(dá)        摘要目的：研究實(shí)驗(yàn)性肝癌的組織發(fā)生過程中甲胎蛋白的表達(dá)，旨在為肝癌診斷奠定形態(tài)學(xué)基礎(chǔ)。方法：Wistar大白鼠喂0.04% 3-Me-DAB不同日程處死，36周處死完，觀察動(dòng)力形態(tài)變化和免疫組化AFP表達(dá)。結(jié)果：實(shí)驗(yàn)早期卵圓細(xì)胞增生，該細(xì)胞是多向分化的干細(xì)胞。它進(jìn)一步分化成過渡細(xì)胞、胚胎性小肝細(xì)胞，后者可呈不典型增生，它們散在或群集，這些細(xì)胞AFP強(qiáng)陽性，群集不典型增生的小肝細(xì)胞形態(tài)上不同于肝細(xì)胞增生結(jié)節(jié)，前者和肝細(xì)

2、胞癌形態(tài)相似。卵圓細(xì)胞向膽管分化形成纖維腺瘤樣增生，其上皮細(xì)胞也見不典型增生。58只實(shí)驗(yàn)動(dòng)物生成26例肝癌，其中肝細(xì)胞癌18例，混合癌8例，癌AFP強(qiáng)陽性，其周圍宿主肝細(xì)胞和實(shí)驗(yàn)晚期無癌動(dòng)物的肝細(xì)胞也見程度不同的AFP陽性表達(dá)。結(jié)論：小肝細(xì)胞和膽管上皮細(xì)胞的不典型增生是肝細(xì)胞癌和膽管癌的癌前病變。關(guān)鍵詞肝腫瘤，試驗(yàn)性；增生；甲胎蛋白類；大鼠分類號R735.7文獻(xiàn)標(biāo)識碼A文章編號1001-7399(1999)03-0224-03 Carcinohistogenesis and expression of alpha fetoprotein in experimental hepatocarcin

3、omaZhang Jizeng(Dept of Pathol, General Hospital of Lanzhou Military Area, Lanzhou730050)ABSTRACTPurposeTo investigate the carcinohistogenesis of hepatocarcinoma and AFP expression in oncogenesis and to lay a morpholoogical foundation for diagnosing and curing hepatocarcinoma. MethodsWistar rats wer

4、e fed on 0.04% 3-Me-DAB and killed according to different dates. All the animals were killed during 36 weeks. The morphologic changes of dynamics and expression of alpha fetoprotein (AFP) were observed with immunohistochemical method. ResultsHyperplasia of oval cells occurred in the early experimemt

5、al stage and the oval cells were multipotential stem cell. They further differentiated into transitional cell and embryoid small liver cell, and the latter may form atypical hyperplasia, scattered or crowded. AFP in these cells appeared to be strongly positive expression. The feature was different b

6、etween dysplastic small hepatocytes with crowd and proliferative nodules of hepatocytes. The former was similar to hepatocytic carcinoma morphologic ally. The oval cells differeatiated towards biliary duct to form fibroadenomatoid structure, and atypical hyperplasia was seen in their epithelial cell

7、s. 26 of 58 experimental animals were cases of hepatocarcinoma, among whom 18 cases of hepatocytic carcinoma and 8 mixed carcinoma were found. The hepatocellular carcinoma showed stuongly AFP positive expression. In the host hepatocytes around cancer and hepatocytes of non-neoplastic animals of late

8、r experimental period also appered to be various degrees of positive expression of the AFP. ConclusionThe precancerosis of hepatocellular carcinoma and bile duct caricnoma is atypical hyperplasia of small hepatocytes and epithelial cells of bile canaliculi.KEY WORDSliver neoplasms, experimental; hyp

9、erplasia; alpha-fetoproteins; rats1973年Anthony等1在烏干達(dá)研究人的肝硬化和肝癌時(shí)首次提出肝細(xì)胞不典型增生是癌前病變。Roncalli等2又把不典型增生的肝細(xì)胞分成大細(xì)胞和小細(xì)胞，認(rèn)為后者對肝細(xì)胞癌的生成關(guān)系密切。1988年徐元鼎等3認(rèn)為肝細(xì)胞癌來自肝細(xì)胞的嗜堿性和嗜酸性增生結(jié)節(jié)。誠然，癌組織發(fā)生是很復(fù)雜的，特別是AFP在肝癌形成中的變化。AFP是確定細(xì)胞分化和腫瘤的標(biāo)記物4。此外，以往對膽管癌的組織發(fā)生以及小膽管上皮細(xì)胞不典型增生與癌發(fā)生的相關(guān)形態(tài)描述也較欠缺。為探討肝癌發(fā)生的動(dòng)力形態(tài)學(xué)，筆者試結(jié)合甲胎蛋白的變化對肝癌的組織發(fā)生進(jìn)行深入研究。1材料

10、與方法Wistar大白鼠喂0.04% 3-Me-DAB混合食物，其只數(shù)與誘癌時(shí)間見表1。在相應(yīng)時(shí)間里取2只正常動(dòng)物肝存檔蠟塊做切片對照。肝標(biāo)本系10%中性福爾馬林固定。AFP免疫組化用ABC法。一抗為兔抗大鼠AFP 1400稀釋；二抗是生物素標(biāo)記的馬抗兔IgG血清1200稀釋；三抗為ABC-HRP(Horseradish Peroxidase)1100稀釋，切片顯色用0.04-H2O2表158只實(shí)驗(yàn)性大白鼠在不同日程內(nèi)誘發(fā)的肝臟病變病變實(shí)驗(yàn)日程(周)<4512132021282936卵圓細(xì)胞增生441過渡細(xì)胞增生43胚胎性小肝細(xì)胞增生668肝細(xì)胞不典型增生5633肝細(xì)胞增生灶或結(jié)節(jié)552

11、0肝假小葉4638小膽管纖維腺瘤樣增生4638肝癌2321處死動(dòng)物只數(shù)446638     表258例實(shí)驗(yàn)性大白鼠肝臟病變中AFP陽性表達(dá)病變nAFP+<"3 (92 bytes)" src="/med/cano/201003/20100318190719247" 16 13><"2 (101 bytes)" src="/med/cano/201003/20100318190719463" 21 13>-陽性率(%)過渡細(xì)胞714271.4胚胎樣小肝細(xì)

12、胞204610100.0不典型增生的肝細(xì)胞4471027100.0肝細(xì)胞增生灶或結(jié)節(jié)301119100.0假小葉內(nèi)的肝細(xì)胞48151016785.4小膽管纖維腺瘤樣增生的上皮細(xì)胞483423918.7癌細(xì)胞*261016100.0     *僅見肝細(xì)胞癌 2結(jié)果1 不典型增生的小肝細(xì)胞(右中下方)和肝癌細(xì)胞(左下方).HE×1002 不典型增生的小肝細(xì)胞AFP(+),而大肝細(xì)胞AFP(-).ABC×2003 肝癌細(xì)胞AFP(+).ABC×4004 小膽管的纖維腺瘤樣增生.HE×2005 小膽管的不典型增生,AFP(

13、+).ABC×20026例肝癌中肝細(xì)胞癌18例；混合癌8例。癌周肝細(xì)胞有不同程度的AFP陽性反應(yīng)。實(shí)驗(yàn)晚期無癌動(dòng)物的肝細(xì)胞亦呈不同程度的AFP陽性反應(yīng)(6)。6 無癌動(dòng)物和肝細(xì)胞AFP(+).ABC×4003討論不典型增生小肝細(xì)胞應(yīng)看作肝細(xì)胞癌的癌前細(xì)胞。它們散在于肝小葉，小葉破壞后的殘留肝細(xì)胞也可形成，因近似胚胎早期的肝細(xì)胞故AFP強(qiáng)陽性反應(yīng)。不典型增生的小肝細(xì)胞群和肝細(xì)胞增生結(jié)節(jié)不同，周圍無壓迫性境界。它們常和肝細(xì)胞癌相鄰，有時(shí)兩者也難以辨認(rèn)，但后者瘤細(xì)胞索形成，其間存有血竇。綜上所述，不典型增生在動(dòng)力形態(tài)的改變過程中有自己的演變過程、形態(tài)特點(diǎn)和存在形成。戴益民等5研究

14、了人體肝癌癌周宿主肝細(xì)胞自己合成AFP。本研究中除肝細(xì)胞癌有AFP陽性表達(dá)外，在實(shí)驗(yàn)晚期無癌動(dòng)物的肝細(xì)胞也有程度不同的AFP陽性表達(dá)。顯然，這些細(xì)胞在致癌因子的作用下雖無形態(tài)上的異型性，但功能上已有返祖改變而處于胚胎狀態(tài)，出現(xiàn)了AFP陽性表達(dá)。肝細(xì)胞癌相連的“正常”肝細(xì)胞索，可視為返祖功能改變的肝細(xì)胞被致癌因子啟動(dòng)失分化后形成的癌突變。實(shí)驗(yàn)晚期無癌動(dòng)物的肝細(xì)胞AFP陽性為臨床非肝癌病人血清AFP水平的增高或提示癌突變的可能性提供了實(shí)驗(yàn)依據(jù)。眾所周知，肝細(xì)胞和膽管細(xì)胞來源于同一胚芽。在實(shí)驗(yàn)性肝癌中Dempo6和Hixson等7指出卵圓細(xì)胞是胚胎性多潛能分化的干細(xì)胞，不言而喻，它既可生成肝細(xì)胞癌也

15、可生成膽管癌和混合性肝癌。明顯的小膽管纖維腺瘤樣增生處卵圓細(xì)胞的消失與它向小膽管分化有關(guān)。不典型增生的小膽管上皮細(xì)胞在間變過程中返祖到與肝細(xì)胞同源的胚胎性質(zhì)時(shí)即可出現(xiàn)AFP陽性表達(dá)，一旦這些細(xì)胞過渡到膽管癌時(shí)其本質(zhì)已不是肝細(xì)胞，故AFP呈陰性反應(yīng)，細(xì)胞角蛋白和上皮細(xì)胞膜抗原呈陽性反應(yīng)，并以此同腺樣結(jié)構(gòu)的肝細(xì)胞癌鑒別。作者簡介：張繼增，男，73歲，教授，主任醫(yī)師。主要從事癌前病變的病理形態(tài)學(xué)研究作者單位：張繼增（蘭州軍區(qū)總醫(yī)院病理科，蘭州730050）參考文獻(xiàn)1，Anthony PP, Vogel CL, Barker LF. Liver cell dysplasia. A premaligna

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17、ein (AFP) expression in clones of MCA-RH7777rat hepatoma: correlation with the occurrence of homogeneously staining regions on chromosome 14. Cancer Res,1995;55(16):361536225，戴益民，倪燦榮. 人體肝癌宿主肝細(xì)胞甲胎蛋白來源的初步探討(附48例免疫組化分析). 中華病理學(xué)雜志，1988；17(4)：2472506，Dempo K, Shisak N, Yoshida Y et al. Immunofluorescent study on a-Fet