倉鼠頰囊袋內鱗狀細胞癌施予化學藥物Cisplatin治療前後細胞週期內各時期之變化－以雷射流體細胞分析儀測定

Squamous Cell Carcinoma of the Pouch of Hamster Treated with Cisplatin-Using Flow Cytometry

洪堅銘(James-Mint Hong) ； 謝天渝(Tien-Yu Shieh) ； 陳中和(Chung-Ho Chen)

《中華民國口腔顎面外科學會雜誌》 8卷1期 (1997/03) Pp. 1-13

https://doi.org/10.7104/CJOMS.199703.0001

摘要

雷射流體細胞分析儀(Laser Flow Cytometry)應用在腫瘤細胞之偵測，與染色體套數之研究已有很多論文發表，並與病理學上組織分類(poor, moderate, well-differention)，與臨床分期(TNM)，已漸被採用來當做預後評估參考之依據。在臨床應用上亦可用來當做化學藥物治療時腫瘤細胞反應的指標，並進而當做治療方針依據。然而有關化學藥物治療隨時間變化，是否會影響到細胞核改變的報告，並不多見。因此，本研究是以DMBA致癌劑定期塗林倉鼠(Hamster)口腔黏膜，致癌後，施予cisplatin化學藥物治療，在注射前、注射後一天、注射後三天、注射後五天隨意選取腫瘤細胞觀蔡其核內細胞週期(cell cycle)各個不同時期，如間期(the interphase, G0+G1)，合成期(synthesis)及後期與分裂期(G2+M)所佔之比例的變化，並進而評估腫瘤細胞對藥物之反應與另一種藥物加入時的適當時機。
倉鼠頰內在定期以0.5% DMBA 塗林在雙側頰囊袋上皮，約八週後即呈現乳突狀之突起，在十至十二週時即有明顯之塊狀突起。本實驗中倉鼠在塗林十六週後其雙側頰內均可見明顯之腫瘤，並切取一小塊送病理檢驗證實為鱗狀細胞癌，並各以三隻有腫瘤細胞之倉鼠與未曾予塗林DMBA之正常頰黏膜做比較，結果顯示正常黏膜之細胞核內S-phase所佔之百分比為8.5%較有腫瘤細胞黏膜的細胞核內S-phase所佔之百分比為26.3%均來得小。進而選取十隻有腫瘤細胞的倉鼠依體重施予cisplatin藥物注射，在注射前，注射後一天、注射後三天、注射後五天隨意選取腫瘤組織，個別以雷射流體細胞分析儀分析其細胞週期內G0+G1，S(synthesis)與G2+M各個時期的百分比。結果顯示在注射後第一天S時期所佔之百分比有明顯降低之趨勢，亦即藥物在第一天有明顯地抑制細胞分化的作用，在第三天S時期有稍微回升。第五天S時期回升地較明顯，此意謂著在目前化學治療中所採取的合併療法(Combined Chemotherapy)在第二種藥物加入的通當時機，以cisplatin注射後隔天加入為宜。至於實際臨床應用上，或人體口腔癌患者接受化學治療前後腫瘤細胞細胞核之變化則有待做更進一步研究。

關鍵字

雷射流體細胞分析儀 ； 化學治療 ； 細胞週期

並列摘要

There are many articles reviewed the studies of the laser Flow cytometry (FCM) for the detectio of DNA polarity and cell kinetics distribution, in combina-tion histologic grading, clinical stage using for as another valuable pro-gnostic indicator. In clinical application, FCM data can be used for the determination of the dose response and the strategy of chemotherapy. However the studies of the cell cycle phase distribution changes during chemotherapy scheduling are rare. In this studies, we induced tumor in hamster pouch mucosa with DMBA, then treated with cisplatin. The cell cycle phase percent-age changes at different time after medication were investigated to evaluate the effects of cisplatin, and provide information on the selection of the best time for giving another drug (s) during combined chemotherapy.
A 0.5% DMBA solution was painted on the pouch mucosa three times every week. After 8 weeks, papillary-like lesions were found. At 10-12 weeks the exophy-tic masses were noted, and at 16 weeks the exophytic masses were larger then the pouch size, and histopathologic diagnosis the squamous cell carcinoma was confirmed. We compared the control normal mucosa of the hamster and the tumor masses, and find that the S-phase average percentage of the normal mucosa is 8.5%, and the tumor is 26.3%. Then these animals were treated with Cisplatin, biopsies were obtained immediately prior to drug administration, 1-day post -ingection, 3-day post-injection, 5-day post -injection. Detection of S-phase% changes using flow cytometry were performed. There was an initial decrease of the fraction of cell in the S phase concomitant with transient depression of DNA synthesis in the 1-day post-injection. The S-phase% is the lowest, in 1 day post- injection suggesting that it may be of the most important period for scheduling combination of other drug (s).