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制首乌含药血清对人乳腺癌T-47D细胞增殖及ER表达的影响
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| 篇名： | 制首乌含药血清对人乳腺癌T-47D细胞增殖及ER表达的影响 |
| TITLE： | |
| 摘要： | 目的：研究制首乌含药血清对人乳腺癌T-47D细胞增殖及雌激素受体（ER）表达的影响，探讨其植物雌激素（PE）样作用。方法：将性未成熟SD大鼠随机分为空白组，戊酸雌二醇（Ev）组（阳性对照，0.12 mg/kg），制首乌低、高剂量组（0.75、3 g/kg，以生药量计）以及制首乌低、高剂量+Ev联用组（剂量同各药单用组），每组10只。空白组大鼠灌胃等体积水，各给药组大鼠灌胃相应药物，早晚各1次，连续4 d。末次给药2 h后采血，制备空白血清和含药血清。将T-47D细胞随机分为空白组，Ev组，制首乌低、高剂量组以及制首乌低、高剂量+Ev联用组，分置于含20％空白或相应含药血清的培养基中培养，采用CCK-8法检测各组细胞的增殖率（PR），采用Western blotting法和逆转录-聚合酶链反应法检测ER-α、ER-β蛋白及其mRNA的表达情况。结果：与空白组比较，各给药组细胞的PR[各给药组（24 h），除制首乌高剂量+Ev联用组外的其余各给药组（48、72 h）]均显著升高；且制首乌高剂量组（72 h）显著高于Ev组，制首乌低剂量组+Ev联用组（72 h）显著高于同剂量制首乌单用组，而制首乌高剂量+Ev联用组（72 h）显著低于同剂量制首乌单用组（P＜0.05或P＜0.01）。Ev组、制首乌高剂量组和制首乌低剂量+Ev联用组细胞中ER-α蛋白，各给药组细胞中ER-α mRNA和ER-β蛋白以及Ev组、制首乌低剂量组和制首乌+Ev联用组细胞中ER-β mRNA的相对表达量均显著升高；Ev组细胞中ER-α蛋白及其mRNA的相对表达量均显著高于制首乌单用和联用组；Ev组细胞中ER-β蛋白及其mRNA的相对表达量均显著低于制首乌低剂量+Ev联用组，但ER-β mRNA的相对表达量显著高于制首乌单用组和制首乌高剂量+Ev联用组；制首乌低剂量+Ev联用组细胞中ER-α、ER-β蛋白及其mRNA以及制首乌高剂量+Ev联用组细胞中ER-β mRNA的相对表达量均显著高于同剂量制首乌单用组，而制首乌高剂量+Ev联用组细胞中ER-α蛋白及其mRNA相对表达量均显著低于同剂量制首乌单用组（P＜0.05或P＜0.01）。结论：制首乌含药血清可促进人乳腺癌T-47D细胞的增殖，并可通过促进ER-α和ER-β蛋白及其mRNA的表达来发挥PE样作用。但上述作用弱于雌激素，且两者联合可能会拮抗雌激素的作用。 |
| ABSTRACT： | OBJECTIVE： To study the effects of processed Polygonum multiflorum containing serum on the proliferation and the expression of estrogen receptor （ER） of human breast cancer T-47D cells， and to investigate its phytoestrogen （PE）-like effect. METHODS： Sexually immature SD rats were randomly divided into estradiol valerate （Ev） group （positive control， 0.12 mg/kg）， processed P. multiflorum low-dose and high-dose groups （0.75， 3 g/kg， by crude drug）， low-dose and high-dose processed P. multiflorum+Ev groups （same dose as single drug group）， with 10 rats in each group. Blank group was given constant volume of water intragastrically， and administration groups were given relevant medicine intragastrically； once day and night， for consecutive 4 days. Two hours after last administration， blank serum and containing serum were prepared. T-47D cells were also randomly divided into blank group， Ev group， low-dose and high-dose processed P. multiflorum groups， low-dose and high-dose processed P. multiflorum+Ev groups， and then were cultured in medium which contained 20％ blank serum or drug containing serum. CCK-8 assay was used to detect proliferation rate （PR）. Western blotting assay and RT-PCR were used to detect the protein and mRNA expression of ER-α and ER-β. RESULTS： Compared with blank group， PR of administration groups [each administration group （24 h）， other administration groups （48， 72 h） except for high-dose processed P. multiflorum+Ev group] were increased significantly； high-dose processed P. multiflorum group （72 h） was significantly higher than Ev group， and low-dose processed P. multiflorum+Ev group （72 h） was significantly higher than the same-dose processed P. multiflorum group； high-dose processed P. multiflorum+Ev group （72 h） was significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. Relative protein expression of ER-α in Ev group， high-dose processed P. multiflorum group and low-dose processed P. multiflorum+Ev group， relative mRNA expression of ER-α and protein expression of ER-β in administration groups， relative mRNA expression of ER-β in Ev group， low-dose processed P. multiflorum group and processed P. multiflorum+Ev groups were all increased significantly. Relative protein and mRNA expression of ER-α in Ev group were significantly higher than processed P. multiflorum groups and combination groups. Relative protein and mRNA expression of ER-β in Ev group were significantly lower than low-dose processed P. multiflorum+Ev group， but relative mRNA expression of ER-β was significantly higher than processed P. multiflorum groups and high-dose processed P. multiflorum+Ev group. Relative protein and mRNA expression of ER-α and ER-β in low-dose processed P. multiflorum+Ev group as well as relative mRNA expression of ER-β in high-dose processed P. multiflorum+Ev group were significantly higher than the same-dose processed P. multiflorum group. Relative protein and mRNA expression of ER-α in high-dose processed P. multiflorum+Ev group were significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. CONCLUSIONS： The processed P. multiflorum containing serum can promote the proliferation of human breast cancer T-47D cells， and play the PE-like role through promoting protein and mRNA expression of ER-α and ER-β. However， the above effects are weaker than estrogen， and the combination of the two may antagonize the effect of estrogen. |
| 期刊： | 2019年第30卷第22期 |
| 作者： | 朱璨，王嫣，李尧锋，田敏，唐文超，杨长福，王和生 |
| AUTHORS： | ZHU Can，WANG Yan，LI Yaofeng，TIAN Min，TANG Wenchao，YANG Changfu，WANG Hesheng |
| 关键字： | 制首乌；含药血清；T-47D细胞；细胞增殖；雌激素受体；植物雌激素样作用 |
| KEYWORDS： | Processed Polygonum multiflorum； Containing serum； T-47D cells； Cell proliferation； Estrogen receptor； Phytoestrogen-like effect |
| 阅读数： | 357 次 |
| 本月下载数： | 7 次 |
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