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1.成都中医药大学 附属医院,代谢性疾病中医药调控四川省重点实验室,成都 610075
2.成都中医药大学 肿瘤研究所,成都 610072
陈宁宁,在读硕士,从事中医肿瘤病证治法与方药研究,E-mail:2416408380@qq.com
严然,硕士,讲师,主治医师,从事中医肿瘤病证治法与方药研究,E-mail:yanran@cdutcm.edu.cn
收稿日期:2022-02-15,
网络出版日期:2022-06-06,
纸质出版日期:2022-12-20
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陈宁宁,蒋义芳,杨懿等.小檗碱与吴茱萸碱联用对结直肠癌细胞迁移、侵袭能力的影响[J].中国实验方剂学杂志,2022,28(24):98-104.
CHEN Ningning,JIANG Yifang,YANG Yi,et al.Effect of Berberine Combined with Evodiamine on Migration and Invasion of Colorectal Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(24):98-104.
陈宁宁,蒋义芳,杨懿等.小檗碱与吴茱萸碱联用对结直肠癌细胞迁移、侵袭能力的影响[J].中国实验方剂学杂志,2022,28(24):98-104. DOI: 10.13422/j.cnki.syfjx.20221529.
CHEN Ningning,JIANG Yifang,YANG Yi,et al.Effect of Berberine Combined with Evodiamine on Migration and Invasion of Colorectal Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(24):98-104. DOI: 10.13422/j.cnki.syfjx.20221529.
目的
2
观察小檗碱与吴茱萸碱联用对结直肠癌HCT116、RKO细胞迁移、侵袭能力的影响,并探究其可能机制。
方法
2
细胞增殖与活性检测-8(CCK-8)法检测小檗碱(30 μmol·L
-1
)、吴茱萸碱(0.8 μmol·L
-1
)及两药联用(30+0.8 μmol·L
-1
)对HCT116及RKO细胞增殖的抑制作用;划痕实验、转移小室(Transwell)实验检测小檗碱、吴茱萸碱及两药联用对HCT116及RKO细胞迁移、侵袭能力的影响;蛋白免疫印迹法(Western blot)检测小檗碱、吴茱萸碱及两药联用对HCT116及RKO细胞中上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)蛋白表达的影响。
结果
2
与空白组比较,小檗碱组和吴茱萸碱组对HCT116及RKO细胞增殖、迁移、侵袭能力无明显抑制作用,两药联用组能显著抑制HCT116及RKO细胞增殖、迁移、侵袭能力(
P
<
0.01);与空白组比较,小檗碱组和吴茱萸碱组对HCT116及RKO细胞中PI3K、N-cadherin、E-cadherin蛋白表达无明显作用,而两药联用组能显著降低HCT116及RKO细胞中PI3K、N-cadherin蛋白表达(
P
<
0.01),升高E-cadherin蛋白表达(
P
<
0.01)。其中,吴茱萸碱组也能显著抑制HCT116及RKO细胞中Akt蛋白表达(
P
<
0.05),但这种抑制作用弱于两药联用组。
结论
2
小檗碱与吴茱萸碱联用能够显著抑制结肠癌HCT116、RKO细胞迁移、侵袭能力,发挥协同增效作用,其作用机制可能与下调PI3K、Akt蛋白表达水平相关。
Objective
2
To observe the effect of berberine combined with evodiamine on the migration and invasion of colorectal cancer HCT116 and RKO cells and to explore the underlying mechanism.
Method
2
cell counting kit-8 (CCK-8) assay was used to examine the proliferation of HCT116 and RKO cells treated by berberine (30 μmol·L
-1
), evodiamine (0.8 μmol·L
-1
), and combination of two (30 μmol·L
-1
+0.8 μmol·L
-1
), respectively. Scratch assay and Transwell assay were employed to detect the migration and invasion of HCT116 and RKO cells treated with berberine, evodiamine, and the combination, separately. In addition, the protein expression of epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt) in HCT116 and RKO cells treated with the berberine, evodiamine, and the combination was respectively measured by Western blot.
Result
2
Compared with the blank group, berberine alone and evodiamine alone had no significant inhibitory effect on the proliferation, migration, and invasion of HCT116 and RKO cells, while the combination showed significant inhibition (
P
<
0.01). Berberine alone and evodiamine alone had no remarkable influence on the expression of PI3K, N-cadherin, and E-cadherin in HCT116 and RKO cells, but the combination significantly reduced the expression of PI3K and N-cadherin (
P
<
0.01) and increased the expression of E-cadherin (
P
<
0.01) in HCT116 and RKO cells. Evodiamine alone also significantly suppressed the expression of Akt protein in HCT116 and RKO cells (
P
<
0.05), but the suppression was weaker than that of the combination.
Conclusion
2
The combination of berberine and evodiamine can significantly inhibit the migration and invasion of colorectal cancer HCT116 and RKO cells and the two show synergy. The mechanism is the likelihood that the combination down-regulates the expression of PI3K and Akt.
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