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1.湖南中医药大学 研究生院,长沙 410208
2.湖南省肿瘤医院,长沙 410013
3.湖南省中医药研究院 附属医院,长沙 410006
焦蕉,在读博士,主治医师,从事中西医结合防治肿瘤疾病研究,E-mail:393739583@qq.com
* 蒋益兰,博士生导师,主任医师,从事中西医结合防治肿瘤疾病研究,E-mail:tianshangren624@163.com
收稿日期:2021-09-14,
网络出版日期:2021-10-15,
纸质出版日期:2021-12-05
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焦蕉,唐麒,蒋益兰等.健脾消癌方通过lncRNA HOTAIR/JAK2/STAT3信号通路抑制结肠癌细胞株HCT116转移的机制[J].中国实验方剂学杂志,2021,27(23):66-71.
JIAO Jiao,TANG Qi,JIANG Yi-lan,et al.Efficacy of Jianpi Xiaoai Prescription in Inhibition of Metastasis of Colon Cancer HCT116 Cells: An Exploration Based on LncRNA HOTAIR/JAK2/STAT3 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(23):66-71.
焦蕉,唐麒,蒋益兰等.健脾消癌方通过lncRNA HOTAIR/JAK2/STAT3信号通路抑制结肠癌细胞株HCT116转移的机制[J].中国实验方剂学杂志,2021,27(23):66-71. DOI: 10.13422/j.cnki.syfjx.20212325.
JIAO Jiao,TANG Qi,JIANG Yi-lan,et al.Efficacy of Jianpi Xiaoai Prescription in Inhibition of Metastasis of Colon Cancer HCT116 Cells: An Exploration Based on LncRNA HOTAIR/JAK2/STAT3 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(23):66-71. DOI: 10.13422/j.cnki.syfjx.20212325.
目的
2
观察健脾消癌方对Hox转录本反义基因间RNA(lncRNA HOTAIR)/ 酪氨酸蛋白激酶2(JAK2)/信号传导及转录激活因子3(STAT3)信号通路的影响,探讨健脾消癌方抑制结肠癌转移的可能作用机制。
方法
2
分析不同细胞株lncRNA HOTAIR的表达情况,采用10%,15%,20%健脾消癌方含药血清作用于HCT116细胞,transwell实验检测健脾消癌方对HCT116细胞的侵袭能力影响,实时荧光定量聚合酶链式反应(Real-time PCR)检测lncRNA HOTAIR,JAK2,STAT3 mRNA的表达,蛋白免疫印迹法(Western blot)检测JAK2,磷酸化STAT3(p-STAT3),STAT3蛋白表达情况。
结果
2
结肠癌细胞株HCT116中lncRNA HOTAIR的表达水平最高,采用此细胞作为观察对象。与空白组比较,健脾消癌方各组的侵袭细胞数均降低(
P
<
0.05,
P
<
0.01);健脾消癌方中剂量组JAK2 mRNA相对表达水平降低(
P
<
0.05);健脾消癌方中、高剂量组的lncRNA HOTAIR,健脾消癌方高剂量组的JAK2 mRNA相对表达水平显著降低(
P
<
0.01)。与空白组比较,健脾消癌方中剂量组的p-STAT3蛋白相对表达水平均降低(
P
<
0.05);健脾消癌方中、高剂量组的JAK2蛋白,健脾消癌方高剂量组的p-STAT3蛋白相对表达水平均明显降低(
P
<
0.01)。
结论
2
健脾消癌方可有效抑制结肠癌细胞转移,其机制可能与抑制lncRNA HOTAIR/JAK2/STAT3信号通路的表达相关。
Objective
2
To observe the effect of Jianpi Xiaoai prescription on long non-coding RNA Hox transcript antisense intergenic RNA (lncRNA HOTAIR)/Janus kinase 2 (JAK2) /signal transducer and activator of transcription 3 (STAT3) signaling pathway and to explore the potential mechanism of Jianpi Xiaoai prescription in suppressing the metastasis of colon cancer.
Method
2
The expression of lncRNA HOTAIR in different cells was analyzed. Following the treatment of HCT116 cells with 10%,15%,and 20% Jianpi Xiaoai prescription -containing serum, the invasive ability of Jianpi Xiaoai prescription on HCT116 cells was assessed by transwell assay. The mRNA expression levels of lncRNA HOTAIR,JAK2,and STAT3 were measured by Real-time polymerase chain reaction (Real-time PCR), and the protein expression levels of JAK2, phosphorylated STAT3 (p-STAT3) and STAT3 by Western blot.
Result
2
The highest expression of lncRNA HOTAIR was detected in HCT116 cells. Compared with the blank group, each Jianpi Xiaoai prescription group exhibited a decreased number of invasive cells (
P
<
0.05,
P
<
0.01). The relative JAK2 mRNA expression in the middle-dose Jianpi Xiaoai prescription group was down-regulated (
P
<
0.05), and the relative lncRNA HOTAIR mRNA expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative JAK2 mRNA expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated (
P
<
0.01). Compared with the blank group,the relative p-STAT3 protein expression was down-regulated in the middle-dose Jianpi Xiaoai prescription group (
P
<
0.05), and the relative JAK2 protein expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative p-STAT3 protein expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated (
P
<
0.01).
Conclusion
2
Jianpi Xiaoai prescription effectively inhibits the metastasis of colon cancer cells, which may be related to the inhibition of lncRNA HOTAIR/JAK2/STAT3 signaling pathway.
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