Pharmacoanalytic Shared Resource (PhASR), Comprehensive Cancer Center, The Ohio State University, Columbus, United States.
Science Exchange, Palo Alto, United States.
Elife. 2020 Oct 19;9:e56651. doi: 10.7554/eLife.56651.
As part of the Reproducibility Project: Cancer Biology, we published a Registered Report (Phelps et al., 2016) that described how we intended to replicate selected experiments from the paper 'Coding-independent regulation of the tumor suppressor PTEN by competing endogenous mRNAs' (Tay et al., 2011). Here, we report the results. We found depletion of putative PTEN competing endogenous mRNAs (ceRNAs) in DU145 cells did not impact 3'UTR regulation using a reporter, while the original study reported decreased activity when , , and were depleted (Figure 3C; Tay et al., 2011). Using the same reporter, we found decreased activity when ceRNA 3'UTRs were overexpressed, while the original study reported increased activity (Figure 3D; Tay et al., 2011). In HCT116 cells, ceRNA depletion resulted in decreased PTEN protein levels, a result similar to the findings reported in the original study (Figure 3G,H; Tay et al., 2011); however, while the original study reported an attenuated ceRNA effect in microRNA deficient (Dicer) HCT116 cells, we observed increased PTEN protein levels. Further, we found depletion of the ceRNAs or did not statistically impact DU145, wild-type HCT116, or Dicer HCT116 cell proliferation. The original study reported increased DU145 and wild-type HCT116 cell proliferation when these ceRNAs were depleted, which was attenuated in the Dicer HCT116 cells (Figure 5B; Tay et al., 2011). Differences between the original study and this replication attempt, such as variance between biological repeats, are factors that might have influenced the results. Finally, we report meta-analyses for each result.
作为《可重复性计划：癌症生物学》的一部分，我们发表了一份注册报告（Phelps 等人，2016 年），其中描述了我们如何打算复制论文《非编码调控肿瘤抑制因子 PTEN 的竞争内源性 mRNAs》（Tay 等人，2011 年）中选定的实验。在这里，我们报告结果。我们发现，在 DU145 细胞中耗尽假定的 PTEN 竞争内源性 mRNAs（ceRNAs）并不影响使用报告基因进行 3'UTR 调节，而原始研究报告说当 、 和 被耗尽时活性降低（图 3C；Tay 等人，2011 年）。使用相同的报告基因，我们发现当 ceRNA 3'UTR 过表达时活性降低，而原始研究报告说活性增加（图 3D；Tay 等人，2011 年）。在 HCT116 细胞中，ceRNA 耗尽导致 PTEN 蛋白水平降低，这一结果与原始研究报告的结果相似（图 3G，H；Tay 等人，2011 年）；然而，虽然原始研究报告说在 miRNA 缺陷（Dicer）HCT116 细胞中 ceRNA 效应减弱，但我们观察到 PTEN 蛋白水平增加。此外，我们发现 ceRNAs 或 的耗尽并不显著影响 DU145、野生型 HCT116 或 Dicer HCT116 细胞的增殖。原始研究报告说，当这些 ceRNAs 被耗尽时，DU145 和野生型 HCT116 细胞的增殖增加，而在 Dicer HCT116 细胞中则减弱（图 5B；Tay 等人，2011 年）。原始研究和本次复制尝试之间的差异，例如生物学重复之间的差异，是可能影响结果的因素。最后，我们报告了每个结果的荟萃分析。
