CAJ | 학술논문

目的：研究中药 QHF 复方对人乳腺癌 MCF -7细胞的体外抗癌效果。方法人乳腺癌 MCF -7细胞，华蟾素：人参皂苷 Rg3：三七总皂苷：香菇多糖=57：1：0.4：7的比例配制中药 QHF 复方，配成5个浓度：QHF1、QHF2、QHF3、QHF4和 QHF5。四甲基偶氮唑盐微量酶反应比色法（MTT 法）检测细胞抑制率、流式细胞仪检测细胞周期、酶联免疫吸附剂测定（ELISA）法检测细胞上清液中血管内皮生长因子（VEGF）含量、Transwell 实验定量观察细胞迁移和侵袭能力。结果 MTT 实验结果显示：不同浓度的 QHF 作用于 MCF -7细胞24、48、72 h 后，随着 QHF 浓度的增加，抑制率不断增加（P <0.01）；且随着时间延长，药物对细胞的抑制率逐渐增加（P <0.01）。24、48、72 h 的中药 QHF 复方 IC50值对应华蟾素浓度分别为214.3、44.8、2.8μl。作用48 h 后，与正常对照组比较，QHF 复方组、盐酸多柔比星组、联合用药组细胞周期位于 G0/ G1期和 S 期的比例增多，位于 G2/ M 期的比例减少（P <0.05）；与联合用药组比较，QHF 复方组、盐酸多柔比星组细胞周期位于 G0/ G1期的比例减少，位于 S 期和 G2/ M 期的比例增多（P<0.05）。ELISA 结果显示：作用48 h 后，与正常对照组比较，QHF1组、QHF2组、QHF3组、QHF4组和 QHF5组上清液中 VEGF 含量均下降（P <0.05）。Transwell 实验结果显示：与正常对照组（127.3±5.7）比较，QHF3组（17.7±5.0）、QHF4组（51.3±2.9）和 QHF5组（63.0±6.6）穿透滤膜的 MCF -7细胞数目减少（P <0.01）。结论中药 QHF 复方在体外可抑制人乳腺癌 MCF -7细胞的增殖和迁移、侵袭能力，并能下调 VEGF 的表达。
목적：연구중약 QHF 복방대인유선암 MCF -7세포적체외항암효과。방법인유선암 MCF -7세포，화섬소：인삼조감 Rg3：삼칠총조감：향고다당=57：1：0.4：7적비례배제중약 QHF 복방，배성5개농도：QHF1、QHF2、QHF3、QHF4화 QHF5。사갑기우담서염미량매반응비색법（MTT 법）검측세포억제솔、류식세포의검측세포주기、매련면역흡부제측정（ELISA）법검측세포상청액중혈관내피생장인자（VEGF）함량、Transwell 실험정량관찰세포천이화침습능력。결과 MTT 실험결과현시：불동농도적 QHF 작용우 MCF -7세포24、48、72 h 후，수착 QHF 농도적증가，억제솔불단증가（P <0.01）；차수착시간연장，약물대세포적억제솔축점증가（P <0.01）。24、48、72 h 적중약 QHF 복방 IC50치대응화섬소농도분별위214.3、44.8、2.8μl。작용48 h 후，여정상대조조비교，QHF 복방조、염산다유비성조、연합용약조세포주기위우 G0/ G1기화 S 기적비례증다，위우 G2/ M 기적비례감소（P <0.05）；여연합용약조비교，QHF 복방조、염산다유비성조세포주기위우 G0/ G1기적비례감소，위우 S 기화 G2/ M 기적비례증다（P<0.05）。ELISA 결과현시：작용48 h 후，여정상대조조비교，QHF1조、QHF2조、QHF3조、QHF4조화 QHF5조상청액중 VEGF 함량균하강（P <0.05）。Transwell 실험결과현시：여정상대조조（127.3±5.7）비교，QHF3조（17.7±5.0）、QHF4조（51.3±2.9）화 QHF5조（63.0±6.6）천투려막적 MCF -7세포수목감소（P <0.01）。결론중약 QHF 복방재체외가억제인유선암 MCF -7세포적증식화천이、침습능력，병능하조 VEGF 적표체。
Objective To investigate the in - vitro anticancer effect of Chinese herbal compound QHF on human breast cancer cell line MCF - 7. Methods Chinese toad bufotoxin,ginsenoside Rg3,panax notoginseng saponins and lentinan were mixed into each other according to the proportion of 57: 1: 0. 4: 7 to prepare into five different concentrations of QHF compound prescriptions:QHF1,QHF2,QHF3,QHF4 and QHF5. The inhibitory effect on MCF - 7 cells were observed by MTT assay, the cell cycle was detected by flow cytometry with PI staining,the protein level of VEGF was assessed by ELISA method,the cells′ migration and invasion were observed quantitatively by transwell assay. Results MTT assay results indicated that:after 24 h,48 h and 72 h,as the concentration of QHF in MCF - 7 cells increased,the inhibition rate also increased(P < 0. 01),and as time increased,the drug′s inhibition rate increased as well(P < 0. 01). The IC50 values of medicine QHF Compound at 24 h,48 h and 72 h corresponded to cinobufotalin concentrations of 214. 3 μl,44. 8 μl and 2. 8 μl respectively. Compared with the control group,the cell cycle were more in G0 / G1 and S phases in QHF compound group,doxorubicin hydrochloride group and the combination therapy group after 48 h,and less in cell ratio of G2 / M phase in the three groups(P < 0. 05);Compared with the combination therapy group,the QHF compound group and the doxorubicin hydrochloride group were less in G0 / G1 phase, more in S and G2 / M phases,showing statistically significant differences(P < 0. 05);ELISA results showed that:after 48 h, compared with the control group,the VEGF levels in supernatants of different concentrations of Chinese herbal compound QHF groups were all decreased(P < 0. 05). Transwell results showed that:compared with the control group(127. 3 ± 5. 7),the number of MCF - 7 cells which penetrated through the filter membrane significantly decreased in QHF3 group(17. 7 ± 5. 0), QHF4 group(51. 3 ± 2. 9)and QHF5 group(63. 0 ± 6. 6)(F = 109. 667,P < 0. 01). Conclusion Chinese herbal com-pound QHF can inhibit the proliferation,migration and invasion of MCF - 7 cells,and it could also down regulate the secretion of VEGF in vitro.