Xu Jianbin, Qian Jun, Zhang Wei, Chen Engeng, Zhang Guolin, Cao Gaoyang, Wang Fei, Shen Xuning, Zhou Wei, Song Zhangfa
Department of Colorectal Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China.
Department of Colorectal Surgery, Xinchang People's Hospital, Xinchang, Zhejiang 312500, P.R. China.
Oncol Rep. 2021 May;45(5). doi: 10.3892/or.2021.8004. Epub 2021 Apr 22.
Following the publication of the above paper, the authors realized that, in their follow‑up experiments, the STAT3 and p65 antibodies they used had already expired prior to the results being published. This affected the confidence that the authors could place in the results published in Figs. 1 and 3E. In addition, the findings were also inconsistent with Fig. 4A and B, potentially causing confusion for the readers. The authors therefore repeated some of these experiments with newly purchased antibodies; they also changed the RT-qPCR results. In the published manuscript, the authors investigated the expression of LYPD8 mRNA expression in tissues from stages I, II, and III whereas in the revised manuscript they have investigated stages II and IV. In addition, the authors have supplemented the manuscript with new transwell assays. The revised figures are presented on the next two pages (Figs 1-4). Repeating these particular experiments has resulted in the following changes being necessary to the text of the published paper (changes are highlighted in bold): i) The fourth sentence in the Abstract, on p. 2389, should read as follows: "The results revealed that the expression of LYPD8 was significantly reduced in the CRC tissue compared with that in precancerous tissue and normal tissue, particularly in stage IV tissue." ('III' has been changed to 'IV'). ii) In the Materials and methods section, "" subsection on p. 2390, the last three sentences should be replaced with the following text: "The histological sections were then stained with the DAB Kit (cat. no. PV‑9000; ZSGB‑BIO, Beijing, China). All sections were observed under a bright‑field microscope (Nikon Corporation, Tokyo, Japan)." iii) In the "" subsection in the right‑hand column, the first four sentences should be revised to the following: "Four CRC cell lines (SW480, SW620, HCT116 and RKO) were used. SW480 (ATCC CCL‑228™, organism, human; tissue, colon; disease, colorectal adenocarcinoma), SW620 (ATCC CCL‑227™, organism, human; tissue, colon; derived from metastatic site, lymph node; disease, colorectal adenocarcinoma), HCT116 (ATCC CCL‑247™, organism, human; tissue, colon; disease, colorectal carcinoma) and RKO (ATCC CRL‑2577™, organism, human; tissue, colon; disease, carcinoma) cells were purchased from the American Type Culture Collection (Manassas, VA, USA). The four cell lines within passages 10 were used in all experiments, and the cell lines were maintained at 37˚C in a humidified incubator containing 5% CO". Also, in line 8 of p. 2391, the cell lines here should be changed to "SW480, SW620 and HCT116 cells", and on line 11, "HT29 cells" should be changed to "RKO cells". iv) In the Results section, the following changes to the text are necessary: In the "" subsection, in the second sentence, "immunofluorescence" should have been written as "immunohistochemistry", and the fourth sentence should have read as follows: "The results of the western blotting showed that the levels of p‑P65/P65 and p‑STAT3/STAT3 gradually increased between stage II and IV (Fig. 1B and C)." Then, the three sentences starting on line 7 on p. 2391 should now read as follows: "Following this, the gene expression levels of LYWPD8 in stage II and IV CRC tissue, precancerous tissue, and normal tissue were assessed using RT‑qPCR analysis (Fig. 2C). Compared with the precancerous tissue and normal tissue, the gene expression of LYPD8 was significantly reduced in stage II and IV tissues. Furthermore, the expression of LYPD8 was reduced in stage IV tissue compared with that in stage II tissue." v) In the subsequent subsection, "", the first sentence should have read as follows: "The plasmid DNA for overexpressing LYPD8 was constructed using the eukaryotic expression vector (pIRES2), as shown in Fig. 3A and B, and the relative expression levels of LYPD8 in the RTO, SW480 HCT116 and SW620 cells were examined by RT-qPCR analysis. vi) In the "" subsection, the penultimate sentence as it appears towards the foot of p. 2392 should now read as follows: "As shown in Fig. 4C and D, a more marked inhibitory effect on cell migration was observed in the LYPD8 OE group compared with that in the control, LYPD8 OE + IL‑6 and LYPD8 OE + TNF‑α groups." vi) In the Discussion, the sentence starting on p. 2394, right‑hand column, line 10 should read as follows: "By contrast, the expression of LYPD8 was significantly reduced in stage II and IV CRC tissues." vii) Finally, some revisions were necessary to the descriptions in the figure legends for Figs. 1, 2 and 4, as follows (only the affected text is included, and the changes are indicated in bold): Figure 1. STAT3 and P65 are activated in colonic tumor tissues from patients. (A) Representative immunohistochemistry images revealing activated STAT3 and P65 in colonic cancer tissue and precancerous tissue. Scale bar, 100 µm. (B) Representative western blotting revealing the expression of p‑P65, P65, p‑STAT3 and STAT3 in stages II and IV colonic tumor tissues. GAPDH was used as a control. (C) Band intensities of western blotting for p‑P65/P65 and p‑STAT3/STAT3 in stage II and IV tissues were analyzed. The data are reported as the mean ± standard deviation of experiments (n=4). P<0.05, phosphorylation levels of STAT3 in stage II tissues vs. in stage IV tissues. Figure 2. Association between IL‑6/TNF‑α and the expression of LYPD8 in colonic tumor tissue, precancerous tissue and normal tissue at different stages. (A) IL‑6 and (B) TNF‑α secretion were analyzed by ELISA in stage II and IV colonic tumor tissue and precancerous tissue. (C) Gene expression of LYPD8 in stage II and IV colonic tumor tissue and precancerous tissue. β‑actin was used as a control. The data are reported as the mean ± standard deviation of experiments (n=6). P<0.01, LYPD8 mRNA expression of normal tissue, precancerous tissue vs. colonic tumor tissue in stage II and IV tissues. Figure 4. Effects of the overexpression of LYPD8 on SW480 cell proliferation and migration. (A) Cell viability of the Control, LYPD, LYPD8 OE + IL‑6 and LYPD8 OE + TNF‑α groups of SW480 cells. (B) SW480 cells were treated with different concentrations (0.5, 1 and 2 M) of niclosamide and different concentrations (5, 15 and 30 M) of JSH‑23, respectively. (C) Numbers of migratory SW480 cells from the Control, LYPD8, LYPD8 OE + IL‑6 and LYPD8 OE + TNF‑α groups. (D) Transwell assay of SW480 cells from the (a) Control, (b) LYPD8 OE, (c) LYPD8 OE + IL‑6 and (d) LYPD8 OE + TNF‑α groups (magnification, ×200). Note that the replacement of the original figures and these revisions made to the text do not drastically alter the overall conclusions reported in the study. The authors are very grateful to the Editor of for allowing them the opportunity to publish this Corrigendum; furthermore, they apologize for any inconvenience caused to the readership of the Journal. [the original articles was published in Oncology Reports 41: 2389‑2395, 2019; DOI: 10.3892/or.2019.7034].
上述论文发表之后,作者发现,在他们的后续实验中,所使用的STAT3和p65抗体在结果发表之前就已经过期。这影响了作者对图1和3E中发表结果的信心。此外,这些结果也与图4A和B不一致,可能会给读者造成困惑。因此,作者使用新购买的抗体重复了部分实验;他们还更改了RT-qPCR结果。在已发表的手稿中,作者研究了I、II和III期组织中LYPD8 mRNA的表达,而在修订后的手稿中,他们研究的是II和IV期。此外,作者还在手稿中补充了新的Transwell实验。修订后的图见接下来的两页(图1-4)。重复这些特定实验后,已发表论文的文本有如下必要更改(更改部分用粗体突出显示):i) 第2389页摘要中的第四句话应改为:“结果显示,与癌前组织和正常组织相比,CRC组织中LYPD8的表达显著降低,尤其是在IV期组织中。”(“III”改为“IV”)。ii) 在材料与方法部分第2390页的“……”子部分,最后三句话应替换为以下内容:“然后用DAB试剂盒(货号PV-9000;北京中杉金桥生物技术有限公司,中国北京)对组织切片进行染色。所有切片均在明场显微镜(尼康公司,日本东京)下观察。”iii) 在右手栏的“……”子部分,前四句话应改为:“使用了四种CRC细胞系(SW480、SW620、HCT116和RKO)。SW480(ATCC CCL-228™,生物,人;组织,结肠;疾病,结直肠癌)、SW620(ATCC CCL-227™,生物,人;组织,结肠;源自转移部位,淋巴结;疾病,结直肠癌)HCT116(ATCC CCL-247™,生物,人;组织,结肠;疾病,结肠癌)和RKO(ATCC CRL-2577™,生物,人;组织,结肠;疾病,癌)细胞购自美国典型培养物保藏中心(美国弗吉尼亚州马纳萨斯)。所有实验均使用传代10代以内的这四种细胞系,细胞系在含5% CO₂的湿度培养箱中于37˚C培养。”此外,在第2391页第8行,此处的细胞系应改为“SW480、SW620和HCT116细胞”,第11行的“HT29细胞”应改为“RKO细胞”。iv) 在结果部分,文本有如下必要更改:在“……”子部分,第二句中的“免疫荧光”应改为“免疫组织化学”,第四句应改为:“蛋白质印迹结果显示,p-P65/P65和p-STAT3/STAT3水平在II期和IV期之间逐渐升高(图1B和C)。”然后,第2391页第7行开始的三句话应改为:“随后,使用RT-qPCR分析评估II期和IV期CRC组织、癌前组织和正常组织中LYWPD8的基因表达水平(图2C)。与癌前组织和正常组织相比,LYPD8的基因表达在II期和IV期组织中显著降低。此外,与II期组织相比,LYPD8在IV期组织中的表达降低。”v) 在随后的“……”子部分,第一句应改为:“如图3A和B所示,使用真核表达载体(pIRES2)构建了用于过表达LYPD8的质粒DNA,并通过RT-qPCR分析检测了RTO、SW480、HCT116和SW620细胞中LYPD8的相对表达水平。vi) 在“……”子部分,第239页接近底部的倒数第二句应改为:“如图4C和D所示,与对照、LYPD8 OE + IL-6和LYPD8 OE + TNF-α组相比,在LYPD8 OE组中观察到对细胞迁移的抑制作用更明显。”vii) 在讨论部分,第2394页右手栏第10行开始的句子应改为:“相比之下,LYPD8的表达在II期和IV期CRC组织中显著降低。”viii) 最后,对图1、2和4的图例描述也进行了一些修订,如下(仅包含受影响的文本,更改部分用粗体表示):图1. STAT3和P65在患者结肠肿瘤组织中被激活。(A) 代表性免疫组织化学图像显示结肠癌组织和癌前组织中激活的STAT3和P65。标尺,100 µm。(B) 代表性蛋白质印迹显示II期和IV期结肠肿瘤组织中p-P65、P65、p-STAT3和STAT3的表达。GAPDH用作对照。(C) 分析II期和IV期组织中p-P65/P65和p-STAT3/STAT3的蛋白质印迹条带强度。数据表示为实验的平均值±标准差(n = 4)。P < 0.05,II期组织与IV期组织中STAT3的磷酸化水平。图2. 不同阶段结肠肿瘤组织、癌前组织和正常组织中IL-6/TNF-α与LYPD8表达的关联。(A) IL-6和(B) TNF-α分泌通过ELISA在II期和IV期结肠肿瘤组织和癌前组织中进行分析。(C) II期和IV期结肠肿瘤组织和癌前组织中LYPD8的基因表达。β-肌动蛋白用作对照。数据表示为实验的平均值±标准差(n = 6)。P < 0.01,II期和IV期组织中正常组织、癌前组织与结肠肿瘤组织的LYPD8 mRNA表达。图4. LYPD8过表达对SW480细胞增殖和迁移的影响。(A) SW480细胞的对照、LYPD、LYPD8 OE + IL-6和LYPD8 OE + TNF-α组的细胞活力。(B) SW480细胞分别用不同浓度(0.5、1和2 μM)的氯硝柳胺和不同浓度(5、15和30 μM)的JSH-23处理。(C) 对照、LYPD8、LYPD8 OE + IL-6和LYPD8 OE + TNF-α组的SW480迁移细胞数量。(D) SW从对照、(a) LYPD8 OE、(c) LYPD8 OE + IL-6和(d) LYPD8 OE + TNF-α组的480细胞的Transwell实验(放大倍数,×200)。请注意,原始图的替换以及对文本的这些修订并没有大幅改变该研究报告的总体结论。作者非常感谢《……》的编辑给予他们发表这份勘误的机会;此外,他们对给该期刊读者带来的不便表示歉意。[原始文章发表于《肿瘤学报道》41: 2389 - 2395, 2019;DOI: 10.3892/or.2019.7034]