Department of Gastrointestinal Oncology, Harbin Medical University Cancer Hospital, Harbin, 150081, Heilongjiang, China.
Lasers Med Sci. 2020 Oct;35(8):1741-1750. doi: 10.1007/s10103-020-02966-8. Epub 2020 Feb 8.
To investigate whether the Warburg effect is a key modulator on the resistance mechanism of photodynamic therapy (PDT). Glycolysis was examined by the test of lactate product and glucose uptake at different post-PDT time points. Cell viability was detected by the CCK-8 assay and cell proliferation was detected by colony formation assay. The expression of glycolysis and related proteins were examined by western blotting. Target gene was silenced by RNAi. In the present study, we assessed the effect of PDT on cancer cell glycolysis. Our team has demonstrated that pyruvate kinase M2 (PKM2), a key speed-limiting enzyme of glycolysis, was significantly overexpressed in patients with esophageal cancer. Our results in the present study showed that PKM2 was downregulated, and lactate product and glucose uptake were inhibited in cells exposed to 5-aminolevulinic acid (5-ALA)-mediated PDT at 4 h after treatment. However, at 24 h after PDT, we observed a substantial increase in PKM2 expression, lactate product, and glucose uptake. Moreover, silencing of PKM2 gene abrogated the upregulatory effect of PDT on glycolysis at late post-PDT period. 2-Deoxy-D-glucose (2-DG) is a recognized chemical inhibitor of glycolysis. The combined treatment of 2-DG and PDT significantly inhibited tumor growth in vitro at 24 h. These results demonstrate that PDT drives the Warburg effect in a time-dependent manner, and PKM2 plays an important role in this progress, which indicated that PKM2 may be a potential molecular target to increase the sensitivity of esophageal cancer cells to PDT.
为了研究瓦博格效应是否是影响光动力疗法(PDT)抵抗机制的关键调节因子。通过不同 PDT 后时间点的乳酸产物和葡萄糖摄取试验来检测糖酵解。通过 CCK-8 测定法检测细胞活力,通过集落形成测定法检测细胞增殖。通过 Western blot 检测糖酵解和相关蛋白的表达。通过 RNAi 沉默靶基因。在本研究中,我们评估了 PDT 对癌细胞糖酵解的影响。我们的团队已经证明,丙酮酸激酶 M2(PKM2),糖酵解的关键限速酶,在食管癌患者中显著过表达。我们本研究的结果表明,在接受 5-氨基酮戊酸(5-ALA)介导的 PDT 治疗后 4 小时,PKM2 下调,细胞中乳酸产物和葡萄糖摄取受到抑制。然而,在 PDT 后 24 小时,我们观察到 PKM2 表达、乳酸产物和葡萄糖摄取显著增加。此外,沉默 PKM2 基因可消除 PDT 对晚期糖酵解的上调作用。2-脱氧-D-葡萄糖(2-DG)是一种公认的糖酵解化学抑制剂。2-DG 和 PDT 的联合治疗在 24 小时时显著抑制了体外肿瘤生长。这些结果表明 PDT 以时间依赖性方式驱动瓦博格效应,PKM2 在这一过程中起重要作用,这表明 PKM2 可能是增加食管癌细胞对 PDT 敏感性的潜在分子靶标。