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目的探讨乳酸菌培养上清液(lactobacilli supernatants,LS)处理乳腺癌细胞对顺铂(cisplatin,DDP)敏感性的影响及其机制。方法采用CCK-8和Western blot检测不同浓度DDP(0、1、2、3、4、5μg/m L)和LS(体积分数分别为1%、5%、10%、20%)对三阴性乳腺癌细胞MDA-MB-231增殖和Piwil2基因表达的影响;瞬时转染Piwil2 siRNA沉默MDA-MB-231细胞中Piwil2基因表达,Western blot检测其沉默效果,CCK-8检测Piwil2基因沉默后对MDA-MB-231细胞增殖的影响;2μg/m L DDP联合不同浓度LS(体积分数分别为1%、5%、10%、20%)作用MDA-MB-231细胞,观察细胞增殖抑制效应和Piwil2蛋白表达的改变。结果高浓度DDP(≥3μg/m L)可抑制MDA-MB-231细胞的增殖(P<0.05),而低浓度DDP(≤2μg/m L)不能有效抑制细胞增殖;DDP以剂量依赖性方式上调Piwil2蛋白的表达;siRNA沉默Piwil2表达可恢复2μg/m L DDP对细胞增殖的抑制。与对照组(乳酸菌培养基,MRS)比较,LS以剂量依赖性方式下调Piwil2蛋白的表达,但并不影响细胞的增殖;与单用2μg/m L DDP比较,2μg/m L DDP联合20%LS处理可显著下调Piwil2蛋白的表达并抑制细胞增殖(P<0.05)。结论三阴性乳腺癌细胞中Piwil2的高表达可导致细胞对顺铂敏感性的降低,乳酸菌培养上清液可通过下调Piwil2基因的表达增强三阴性乳腺癌细胞对顺铂的敏感性。
Objective To investigate the effect of lactobacilli supernatants (LS) on the sensitivity of breast cancer cells to cisplatin (DDP) and its mechanism. Methods CCK-8 and Western blot were used to detect the effect of different concentrations of DDP (0, 1, 2, 3, 4, 5μg / mL) and LS (1%, 5%, 10%, 20% The expression of Piwil2 gene in MDA-MB-231 cells was transiently transfected with Piwil2 siRNA. The silencing effect of Piwil2 gene was detected by Western blot. The effect of 2μg / ml DDP on MDA-MB-231 cells with different concentrations of LS (1%, 5%, 10%, 20% Changes in protein expression. Results DDP (≥3μg / m L) could inhibit the proliferation of MDA-MB-231 cells (P <0.05), while DDP (≤2μg / mL) did not inhibit cell proliferation effectively. Upregulated the expression of Piwil2 protein; siRNA silencing Piwil2 expression restored the inhibition of cell proliferation by 2μg / m L DDP. Compared with the control group (lactic acid bacteria medium, MRS), LS down-regulated the expression of Piwil2 protein in a dose-dependent manner, but did not affect the cell proliferation. Compared with 2μg / m L DDP alone, LS treatment significantly down-regulated Piwil2 protein expression and inhibited cell proliferation (P <0.05). Conclusion The high expression of Piwil2 in triple-negative breast cancer cells can lead to the decrease of cisplatin sensitivity. Lactobacillus culture supernatant can enhance the sensitivity of triple-negative breast cancer cells to cisplatin by down-regulating the expression of Piwil2 gene.