| 陈吉柏,范长玲,张浩.薯莨提取物通过 KTN1反义 RNA 1对食管癌细胞生物行为的影响[J].安徽医药,2022,26(6):1084-1088. |
| 薯莨提取物通过 KTN1反义 RNA 1对食管癌细胞生物行为的影响 |
| Effect of Dioscorea cirrhosa extract on biological behavior of esophageal cancer cells through KTN1-AS1 |
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| DOI：10.3969/j.issn.1009-6469.2022.06.005 |
| 中文关键词: 薯蓣属 薯莨提取物 KTN1反义 RNA 1 食管肿瘤 增殖 迁移 侵袭 |
| 英文关键词: Dioscorea Dioscorea cirrhosa extract KTN1-AS1 Esophageal neoplasms Proliferation Migration Invasion |
| 基金项目:海南省卫生和计划生育委员会科研项目( 1605032027A2001) |
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| 中文摘要: |
| 目的研究薯莨提取物是否影响食管癌细胞的生物学行为。方法自 2019年 1月至 2020年 1月，采用( 10、20和 30 mg/L)浓度的薯莨提取物处理食管癌 Eca109细胞，分别记为薯莨 -L组、薯莨 -M组、薯莨 -H组，另以未加薯莨提取物的 Eca109细胞作为对照组。细胞计数试剂盒 8(CCK-8)测定细胞增殖，克隆形成实验分析细胞克隆能力，蛋白质印迹法检测周期素依赖激酶抑制剂 p21(P21)、上皮钙黏素( E-cadherin)、基质金属蛋白酶 -2(MMP-2)蛋白表达， Transwell分析细胞迁移、侵袭，实时荧光定量逆转录聚合酶链反应( qRT-PCR)检测长链非编码 RNA(lncRNA)KTN1反义 RNA 1(KTN1-AS1)表达。在 Eca109细胞中转染 KTN1-AS1小干扰 RNA(si-KTN1-AS1)，或转染 KTN1-AS1过表达质粒( pcDNA3.1-KTN1-AS1)并使用薯莨提取物处理，观察其对细胞增殖、迁移和侵袭的影响。结果与对照组相比，薯莨 -L组、薯莨 -M组、薯莨 -H组 Eca109细胞的抑制率［( 1.06±0.21)%比( 19.25±1.68)%比( 38.57±3.69)%比( 62.14±6.06)%］、 P21、E-cadherin蛋白水平明显提高，细胞的克隆形成数、迁移细胞数［( 142.00±11.59)比( 121.00±11.07)比( 94.00±9.26)比( 73.00±7.15)］、侵袭细胞数［( 81.00±8.03)比( 67.00±6.21)比( 50.00±5.04)比(37.00±3.56)］、 MMP-2蛋白表达量、 KTN1-AS1表达量显著减少( P<0.05)均呈浓度依赖性。抑制 KTN1-AS1明显增加 Eca109细胞的抑制率［(1.04±0.17)%比( 47.81±4.55)%］、 E-cadherin、P21蛋白表显著降低迁移细胞数［(144.00±13.52)比达量，(85.00±8.51)］、侵袭细胞数［(80.00±8.11)比( 47.00±4.36)］、克隆形成数和 MMP-2蛋白水平( P<0.05)。过表达 KTN1-AS1能减弱薯莨提取物对食管癌细胞增殖、迁移、侵袭的抑制作用。结论薯莨提取物通过下调食管癌细胞中 KTN1-AS1的表达，发挥抑制细胞增殖、迁移和侵袭的作用。 |
| 英文摘要: |
| Objective To investigate whether the extract of Dioscorea cirrhosa extract affects the biological behavior of esophageal cancer cells through long non-coding RNA (lncRNA) KTN1 antisense RNA 1 (KTN1-AS1). Methods From January 2019 to January 2020, esophageal cancer Eca109 cells were treated with (10, 20, and 30 mg/L) Dioscorea cirrhosa extract, which were designated as Di? oscorea cirrhosa-L group, Dioscorea cirrhosa-M group and Dioscorea cirrhosa-H group, respectively, and Eca109 cells without Dioscorea cirrhosa extract were used as control group. Cell proliferation was measured by Cell Counting Kit-8 (CCK-8), the cloning formation experiment analyzed the cloning ability of the cells. Western blotting was used to detect the protein expression of P21, E-cadherin and matrix metalloproteinase-2 (MMP-2), Transwell analyzed cell migration and invasion, and real-time fluorescence quantitative PCR (qRT-PCR) detected KTN1-AS1 expression. Eca109 cells were transfected with KTN1-AS1 small interfering RNA (si-KTN1-AS1), or transfected with KTN1-AS1 overexpressed plasmid (pcDNA3.1-KTN1-AS1) and treated with Dioscorea cirrhosa extract to observe the effects on cell proliferation, migration, and invasion. Results Compared with the control group, the inhibition rate [(1.06±0.21) % vs. (19.25± 1.68) % vs. (38.57±3.69) % vs. (62.14±6.06) %] of Eca109 cells in Dioscorea cirrhosa-L group, Dioscorea cirrhosa-M group and Di? oscorea cirrhosa-H group, P21, and E-cadherin protein levels were significantly increased, and the number of colony formation, number of migrating cells [(142.00±11.59) vs. (121.00±11.07) vs. (94.00±9.26) vs. (73.00±7.15)], number of invading cells [(81.00±8.03) vs. (67.00±6.21) vs. (50.00±5.04) vs. (37.00±3.56)], MMP-2 protein expression levels, and KTN1-AS1 expression were evidently reduced (P <0.05), with concentration dependence. Inhibition of KTN1-AS1 greatly increased the inhibition rate [(1.04±0.17) % vs. (47.81±4.55) %] of Eca109 cells, as well as the expression of E-cadherin, and P21 protein, and obviously reduced the number of migrating cells [(144.00±13.52) vs. (85.00±8.51)], invading cells [(80.00±8.11) vs. (47.00±4.36)], colony formation, and MMP-2 protein levels (P< 0.05). Overexpression of KTN1-AS1 can reduce the inhibitory effect of Dioscorea cirrhosa extract on the proliferation, migration and invasion of esophageal cancer cells. Conclusion Dioscorea cirrhosa extract can inhibit cell proliferation, migration and invasion by down-regulating the expression of KTN1-AS1 in esophageal cancer cells. |
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