现代肿瘤医学
現代腫瘤醫學
현대종류의학
JOURNAL OF MODERN ONCOLOGY
2014年
5期
1049-1053
,共5页
乳腺癌%基质金属蛋白酶%免疫组织化学%明胶酶谱%转移
乳腺癌%基質金屬蛋白酶%免疫組織化學%明膠酶譜%轉移
유선암%기질금속단백매%면역조직화학%명효매보%전이
breast cancer%MMP%immunohistochemistry%gelatin zymogram%metastasis
目的:探讨乳腺癌 MMP -2、MMP -9表达的临床病理联系及直接阻断 MMP -2、MMP -9对乳腺癌细胞增殖、侵袭及细胞周期的影响。方法:应用免疫组织化学方法检测48例乳腺癌 MMP -2、MMP -9的表达。明胶酶谱法检测乳腺癌 MCF -7细胞 MMP -2、MMP -9的表达。MCF -7细胞接种于预铺 BME 的 Tran-swell 小室上室,与 MMP -2、MMP -9的阻断剂 CTT 共孵育16h,观察阻断 MMP -2、MMP -9对 MCF -7细胞侵袭能力的影响。MCF -7细胞与 CTT 共孵育12h 后,MTT 法检测阻断 MMP -2、MMP -9对细胞增殖能力的影响。MCF -7细胞与 CTT 共孵育24h 后,PI 染色流式法检测阻断 MMP -2、MMP -9对细胞凋亡和细胞周期的影响。结果:48例乳腺癌组织有 MMP -2表达者45例(93.75%),MMP -9表达者38例(79.17%),MMP-2和 MMP -9在发生淋巴结转移组明显高表达(P ﹤0.05);组织学分级越高,MMP -2和 MMP -9的表达越显著(P ﹤0.05)。明胶酶谱法检测:在 MCF -7培养上清中检测到 MMP -2、MMP -9的表达。在浓度为100μg/ ml 和200μg/ ml 时 CTT 对 MCF -7细胞的侵袭抑制率达到39.5%和61.9%。在终浓度为50、100、200μg/ ml 时,CTT 对 MCF -7细胞的增殖率均无明显下降(P ﹥0.05)。CTT 处理组 MCF -7细胞经流式法检测未见凋亡峰,细胞周期各期未见明显阻滞(P ﹥0.05)。结论:MMP -2、MMP -9与乳腺癌的淋巴结转移密切相关,阻断 MMP -2、MMP -9可有效抑制乳腺癌的浸润和转移。
目的:探討乳腺癌 MMP -2、MMP -9錶達的臨床病理聯繫及直接阻斷 MMP -2、MMP -9對乳腺癌細胞增殖、侵襲及細胞週期的影響。方法:應用免疫組織化學方法檢測48例乳腺癌 MMP -2、MMP -9的錶達。明膠酶譜法檢測乳腺癌 MCF -7細胞 MMP -2、MMP -9的錶達。MCF -7細胞接種于預鋪 BME 的 Tran-swell 小室上室,與 MMP -2、MMP -9的阻斷劑 CTT 共孵育16h,觀察阻斷 MMP -2、MMP -9對 MCF -7細胞侵襲能力的影響。MCF -7細胞與 CTT 共孵育12h 後,MTT 法檢測阻斷 MMP -2、MMP -9對細胞增殖能力的影響。MCF -7細胞與 CTT 共孵育24h 後,PI 染色流式法檢測阻斷 MMP -2、MMP -9對細胞凋亡和細胞週期的影響。結果:48例乳腺癌組織有 MMP -2錶達者45例(93.75%),MMP -9錶達者38例(79.17%),MMP-2和 MMP -9在髮生淋巴結轉移組明顯高錶達(P ﹤0.05);組織學分級越高,MMP -2和 MMP -9的錶達越顯著(P ﹤0.05)。明膠酶譜法檢測:在 MCF -7培養上清中檢測到 MMP -2、MMP -9的錶達。在濃度為100μg/ ml 和200μg/ ml 時 CTT 對 MCF -7細胞的侵襲抑製率達到39.5%和61.9%。在終濃度為50、100、200μg/ ml 時,CTT 對 MCF -7細胞的增殖率均無明顯下降(P ﹥0.05)。CTT 處理組 MCF -7細胞經流式法檢測未見凋亡峰,細胞週期各期未見明顯阻滯(P ﹥0.05)。結論:MMP -2、MMP -9與乳腺癌的淋巴結轉移密切相關,阻斷 MMP -2、MMP -9可有效抑製乳腺癌的浸潤和轉移。
목적:탐토유선암 MMP -2、MMP -9표체적림상병리련계급직접조단 MMP -2、MMP -9대유선암세포증식、침습급세포주기적영향。방법:응용면역조직화학방법검측48례유선암 MMP -2、MMP -9적표체。명효매보법검측유선암 MCF -7세포 MMP -2、MMP -9적표체。MCF -7세포접충우예포 BME 적 Tran-swell 소실상실,여 MMP -2、MMP -9적조단제 CTT 공부육16h,관찰조단 MMP -2、MMP -9대 MCF -7세포침습능력적영향。MCF -7세포여 CTT 공부육12h 후,MTT 법검측조단 MMP -2、MMP -9대세포증식능력적영향。MCF -7세포여 CTT 공부육24h 후,PI 염색류식법검측조단 MMP -2、MMP -9대세포조망화세포주기적영향。결과:48례유선암조직유 MMP -2표체자45례(93.75%),MMP -9표체자38례(79.17%),MMP-2화 MMP -9재발생림파결전이조명현고표체(P ﹤0.05);조직학분급월고,MMP -2화 MMP -9적표체월현저(P ﹤0.05)。명효매보법검측:재 MCF -7배양상청중검측도 MMP -2、MMP -9적표체。재농도위100μg/ ml 화200μg/ ml 시 CTT 대 MCF -7세포적침습억제솔체도39.5%화61.9%。재종농도위50、100、200μg/ ml 시,CTT 대 MCF -7세포적증식솔균무명현하강(P ﹥0.05)。CTT 처리조 MCF -7세포경류식법검측미견조망봉,세포주기각기미견명현조체(P ﹥0.05)。결론:MMP -2、MMP -9여유선암적림파결전이밀절상관,조단 MMP -2、MMP -9가유효억제유선암적침윤화전이。
Objective:To study the clinicoPathological significance of MMP-2/-9 exPressions in breast cancer, and the blocking MMP-2/-9 on breast cancer cells. Methods:The exPressions of MMP-2/-9 were examined in 48 cases of breast cancer by immunohistochemistry. The exPressions of MMP-2/-9 in MCF-7 cells were detected by means of gelatin zymogram. MCF-7 cells were incubated with different concentrations of CTT(MMP-2/-9 blocker)in transwell Precoated with BME for 16h,the invasion of MCF-7 cells was detected. MCF-7 cells were in-cubated with different concentrations of CTT in 96 - microwell Polystyrene Plates for 12h,the Proliferation of MCF-7 cells was detected by means of MTT. Incubated with 200μg/ ml of CTT for 24h,aPoPtosis and cell cycle of MCF-7 cells were detected by means of PI staining flow cytometry. Results:The result of immunohistochemistry:Among 48 breast cancer examPles,there were 45 cases exPressing MMP - 2(93. 75% )and 38 cases exPressing MMP - 9 (79. 17% ). The exPressions of MMP-2/-9 in breast cancer with lymPh node metastasis grouP was higher than no metastasis grouP(P ﹤ 0. 05). The result of gelatin zymogram:MMP-2/-9 could be detected in MCF-7 cell cul-ture suPernatant. Cultured with 100μg/ ml and 200μg/ ml of PePtide CTT,the inhibition ratio of cellular invasion of MCF-7 were 39. 5% and 61. 9% . At all concentrations,the Proliferation ratio of MCF-7 had no significant differ-ence between CTT and blank grouP(P ﹥ 0. 05). The result of flow cytometry showed that no aPoPtotic Peak could be found in MCF-7 cells treated with CTT,and the whole cell cycle had no significant difference between CTT and blank grouP(P ﹥ 0. 05). Conclusion:In breast cancer,the excessive exPressions of MMP-2/-9 may indicate axil-lary lymPh node metastasis,blocking the function of MMP-2/-9 can inhibit the metastasis of breast cancer.