西黄丸醇提液激活法尼酯受体的抗乳腺癌作用研究
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摘要
目的研究西黄丸醇提液抑制荷4T1小鼠乳腺癌肿瘤的生长及作用机制。方法建立荷4T1小鼠乳腺癌模型,以低、中、高剂量(0.39、0.78、1.95 g/kg)ig给予西黄丸醇提液2周,分离肿瘤组织,称质量并计算抑瘤率;体外培养4T1乳腺癌细胞株,加入低、中、高浓度(1.25、2.50、5.00μg/mL)的西黄丸醇提液作用48 h,CCK-8法检测细胞增殖抑制率;TUNEL染色检测细胞凋亡;实时荧光定量PCR(qRT-PCR)法检测4T1细胞中FXR mRNA表达,Western Blotting检测法尼酯受体(FXR)蛋白表达。结果与模型组比较,西黄丸醇提液低、中、高剂量组荷瘤质量均明显下降(P<0.05),且呈剂量相关性;CCK-8结果显示,作用48 h,4T1细胞的增殖抑制率随西黄丸醇提液浓度的升高而增加,高浓度组可达43.13%;TUNEL荧光染色结果显示,与对照组比较,西黄丸醇提液低、中、高浓度组4T1细胞凋亡数量显著增加(P<0.05);qRT-PCR、Western Blotting结果显示,4T1细胞中FXR mRNA、蛋白表达水平随西黄丸醇提液浓度的升高显著上调,且低、中、高浓度组差异均具有统计学意义(P<0.05)。结论西黄丸可能通过激活FXR受体促进肿瘤细胞凋亡从而抑制肿瘤生长。
Objective To study the effect of alcohol extract of Xihuang Pill on the growth of 4T1 mouse breast cancer and its possible mechanism. Methods We established a 4T1 mouse breast cancer model. Model mice were administered with alcohol extract of Xihuang Pill at low, medium and high doses for two weeks. Tumor tissues were then removed and weighed, and the tumor inhibition rate was calculated. The 4T1 breast cancer cell line was cultured in vitro. The ethanol extract of Xihuang pill with low,medium and high concentration(1.25, 2.50, 5.00 μg/mL) were added for 48 hours. Tumor cell activity was detected by CCK-8. The apoptosis of tumor cells was detected by TUNEL staining. The mRNA expression of farnesoid X receptor(FXR) in tumor tissue was detected by qRT-PCR and their protein expression levels were detected by Western Blotting. Results Compared with model group,tumor volumes and tumor weights in the alcohol extract of Xihuang Pill groups decreased significantly with increasing doses of Xihuang Pill, there were significant differences among low, medium and high concentration groups(P < 0.05). CCK-8 results showed that the inhibition rate of tumor cells proliferation was increased with the increase of dose of alcohol extract of Xihuang Pill at 48 h, and that of the high dose group reached 43.13%. TUNEL staining showed that the number of apoptotic tumor cells increased with increasing doses of alcohol extract of Xihuang Pill. qRT-PCR and Western Blotting showed that mRNA and protein expression of FXR in tumor cells increased with increasing doses of alcohol extract of Xihuang pill. There were significant differences among low, medium and high concentration groups(P < 0.05). Conclusion Xihuang Pill might promote tumor cell apoptosis and further inhibit the tumor growth of 4T1 mouse breast cancer. The mechanism may be related to upregulation of FXR.
Objective To study the effect of alcohol extract of Xihuang Pill on the growth of 4T1 mouse breast cancer and its possible mechanism. Methods We established a 4T1 mouse breast cancer model. Model mice were administered with alcohol extract of Xihuang Pill at low, medium and high doses for two weeks. Tumor tissues were then removed and weighed, and the tumor inhibition rate was calculated. The 4T1 breast cancer cell line was cultured in vitro. The ethanol extract of Xihuang pill with low,medium and high concentration(1.25, 2.50, 5.00 μg/mL) were added for 48 hours. Tumor cell activity was detected by CCK-8. The apoptosis of tumor cells was detected by TUNEL staining. The mRNA expression of farnesoid X receptor(FXR) in tumor tissue was detected by qRT-PCR and their protein expression levels were detected by Western Blotting. Results Compared with model group,tumor volumes and tumor weights in the alcohol extract of Xihuang Pill groups decreased significantly with increasing doses of Xihuang Pill, there were significant differences among low, medium and high concentration groups(P < 0.05). CCK-8 results showed that the inhibition rate of tumor cells proliferation was increased with the increase of dose of alcohol extract of Xihuang Pill at 48 h, and that of the high dose group reached 43.13%. TUNEL staining showed that the number of apoptotic tumor cells increased with increasing doses of alcohol extract of Xihuang Pill. qRT-PCR and Western Blotting showed that mRNA and protein expression of FXR in tumor cells increased with increasing doses of alcohol extract of Xihuang pill. There were significant differences among low, medium and high concentration groups(P < 0.05). Conclusion Xihuang Pill might promote tumor cell apoptosis and further inhibit the tumor growth of 4T1 mouse breast cancer. The mechanism may be related to upregulation of FXR.
引文
[1]陈万青,郑荣寿.中国女性乳腺癌发病死亡和生存状况[J].中国肿瘤临床, 2015, 42(13):668-674.
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[7]马杰,王一尧,杨伟,等.西黄丸抗肿瘤作用及其免疫清除功能的实验研究[J].中国中药杂志, 2014, 39(8):1499-1501.
[8]金娟,李志鸿.西黄丸联合化疗治疗乳腺癌30例[J].中华中医药杂志, 2010, 25(5):715-716.
[9]吴超.西黄丸抗肿瘤活性组分筛选及与西黄滴丸药效学和化学成分的比较研究[D].济南:山东中医药大学, 2014.
[10]马杰,杨伟,关硕,等.西黄丸乙醇提取物抗肿瘤作用及其免疫学机制实验研究[J].辽宁中医药大学学报, 2014,16(4):43-45.
[11]王海燕,田菲.西黄丸联合介入化疗治疗中晚期恶性肿瘤26例近期疗效观察[J].云南中医中药杂志, 2012, 33(3):27-28.
[12]沈雁鸣.西黄丸联合西医治疗乳腺癌随机平行对照研究[J].实用中医内科杂志, 2014, 28(3):127-128.
[13] Zimber A, Gespach C. Bile acids and derivatives, their nuclear receptors FXR, PXR and ligands:role in health and disease and their therapeutic potential[J]. Anticancer Agents Med Chem, 2008, 8(5):540-563.
[14] Swales K E, Korbonits M, Carpenter R, et al. The farnesoid X receptor is expressed in breast cancer and regulates apoptosis and aromatase expression[J]. Cancer Res, 2006, 66(20):10120-10126.
[2] Fan L, Strasser-Weippl K, Li J J, et al. Breast cancer in China[J]. Lancet Oncol, 2014, 15(7):279-289.
[3] Journe F, Durbecq V, Chaboteaux C, et al. Association between farnesoid X receptor expression and cell proliferation in estrogen receptor-positive luminal-like breast cancer from postmenopausal patients[J]. Breast Cancer Res Treat, 2009, 115(3):523-535.
[4] Swales K E, Korbonits M, Carpenter R, et al. The farnesoid X receptor is expressed in breast cancer and regulates apoptosis and aromatase expression[J]. Cancer Res, 2006, 66(20):10120-10126.
[5] Koutsounas I, Giaginis C, Theocharis S. Farnesoid X Receptor(FXR)from normal to malignant state[J].Histol Histopathol, 2012, 27(7):835-853.
[6]周杰,商雪莹,佟玲,等.西黄丸HPLC-ELSD指纹图谱研究[J].中草药, 2017, 48(11):2219-2224.
[7]马杰,王一尧,杨伟,等.西黄丸抗肿瘤作用及其免疫清除功能的实验研究[J].中国中药杂志, 2014, 39(8):1499-1501.
[8]金娟,李志鸿.西黄丸联合化疗治疗乳腺癌30例[J].中华中医药杂志, 2010, 25(5):715-716.
[9]吴超.西黄丸抗肿瘤活性组分筛选及与西黄滴丸药效学和化学成分的比较研究[D].济南:山东中医药大学, 2014.
[10]马杰,杨伟,关硕,等.西黄丸乙醇提取物抗肿瘤作用及其免疫学机制实验研究[J].辽宁中医药大学学报, 2014,16(4):43-45.
[11]王海燕,田菲.西黄丸联合介入化疗治疗中晚期恶性肿瘤26例近期疗效观察[J].云南中医中药杂志, 2012, 33(3):27-28.
[12]沈雁鸣.西黄丸联合西医治疗乳腺癌随机平行对照研究[J].实用中医内科杂志, 2014, 28(3):127-128.
[13] Zimber A, Gespach C. Bile acids and derivatives, their nuclear receptors FXR, PXR and ligands:role in health and disease and their therapeutic potential[J]. Anticancer Agents Med Chem, 2008, 8(5):540-563.
[14] Swales K E, Korbonits M, Carpenter R, et al. The farnesoid X receptor is expressed in breast cancer and regulates apoptosis and aromatase expression[J]. Cancer Res, 2006, 66(20):10120-10126.