中国医大讲座

1、如何解析回答如何解析回答SCI论文审稿人的意见论文审稿人的意见 博士研究生：张俊毅博士研究生：张俊毅导导 师：王恩华师：王恩华论文题目论文题目-catenin 和和p120ctn在胞浆中通过与在胞浆中通过与E-cadherin非竞争性结合促进非小细胞肺癌的非竞争性结合促进非小细胞肺癌的恶性表型恶性表型背景材料背景材料黏附分子黏附分子Catenin家族中的家族中的p120ctn和和-catenin与肺癌侵袭、转移与肺癌侵袭、转移的关系已有报道，但该家族的另一成员的关系已有报道，但该家族的另一成员-catenin在肺癌组织中在肺癌组织中的表达情况及临床意义尚不清楚。的表达情况及临床意义尚不清楚。

2、文献报道，前列腺癌中文献报道，前列腺癌中-catenin的转录和蛋白水平增加；宫颈癌的转录和蛋白水平增加；宫颈癌和膀胱癌中存在和膀胱癌中存在-catenin基因的扩增基因的扩增 。与。与p120ctn一样，一样，-catenin即可结合即可结合E-cadherin，入核后又可结合，入核后又可结合Kaiso，均提示，均提示-catenin在肿瘤的恶性表型中可能同样扮演重要角色。在肿瘤的恶性表型中可能同样扮演重要角色。 -catenin可通过改变可通过改变small GTPases活性而促进神经细胞突起的活性而促进神经细胞突起的形成，而突起的形成和数量也是细胞恶性表型的形态学标志，形成，而突起的形

3、成和数量也是细胞恶性表型的形态学标志，那么那么-catenin是否通过影响是否通过影响small GTPases活性而对肺癌细胞的活性而对肺癌细胞的恶性表型起到促进作用恶性表型起到促进作用 ？-catenin和和p120ctn都只结合都只结合E-cadherin上的上的JMD位点，那么位点，那么-catenin和和p120ctn之间存在竞争性吗之间存在竞争性吗？-catenin对肺癌细胞生物学行为有何影响对肺癌细胞生物学行为有何影响？需要解决的问题需要解决的问题 结果结果1：-catenin在肺癌组织中表达显著增加在肺癌组织中表达显著增加气道上皮气道上皮黏膜下腺体黏膜下腺体腺腺 癌癌鳞鳞 癌癌

4、结果结果1：-cat, p120ctn, E-cad的表达与临床病理因素的关系的表达与临床病理因素的关系临床病理因素临床病理因素n-catenin positive expressionPp120ctn abnormal expressionPE-cad abnormal expressionP年龄年龄(岁岁) 58 58 526332400.829 45540.89943480.393性别性别 男男 女女615437350.645 52470.78246450.297组织类型组织类型 鳞癌鳞癌 腺癌腺癌 546128440.025 43560.06040510.209分化分化 高中高中 低低

5、813447250.117 66330.02860310.039pTNM分期分期 期期 期期 57583042 0.028 45540.0283952 0.005淋巴结转移淋巴结转移 无无 有有33821557 0.016 24750.00921700.009 结果结果2：-catenin在在LN转移灶中的表达强于原发灶转移灶中的表达强于原发灶原原 发发 灶灶转转 移移 灶灶结果结果2：-catenin在原发灶和在原发灶和LN转移灶中的表达转移灶中的表达 （50% vs. 88% ） 原发灶原发灶P LN转移灶转移灶 600.022 1925结果结果3：-catenin阳性表达患者的生存时间缩

6、短阳性表达患者的生存时间缩短 （Log-Rank检验，检验，P=0.019 ） 结果结果4：-catenin的的mRNA和蛋白在肺癌和蛋白在肺癌 组织中表达增加（组织中表达增加（P均均0.05） 结果结果5：过表达（或干扰）过表达（或干扰）-catenin与与p120ctn后在蛋后在蛋 白水平的验证白水平的验证 结果结果6：-catenin和和p120ctn均可与均可与E-cadherin结合，结合， 但二者并不存在竞争关系但二者并不存在竞争关系 结果结果7：E-cadherin、-catenin和和p120ctn在肺癌细在肺癌细 胞系中主要定位于胞浆胞系中主要定位于胞浆 结果结果8：-cat

7、enin可下调可下调RhoA活性，上调活性，上调Cdc42和和 Rac1的活性的活性 结果结果9：-catenin可改变肺癌细胞周期可改变肺癌细胞周期 结果结果10： -catenin能促进肺癌细胞增殖能促进肺癌细胞增殖 结果结果11：-catenin能改变肺癌细胞骨架的排列能改变肺癌细胞骨架的排列 （F-actin荧光染色）荧光染色）结果结果12：-catenin能促进肺癌细胞的侵袭能促进肺癌细胞的侵袭结论结论-catenin在肺癌组织中高表达并与患者的不良预后相关。在肺癌组织中高表达并与患者的不良预后相关。-catenin和和p120ctn均可与均可与E-cadherin在肺癌细胞胞浆中结

8、在肺癌细胞胞浆中结合，但两者不存在竞争关系。合，但两者不存在竞争关系。-catenin通过降低通过降低RhoA活性、升高活性、升高Cdc42和和Rac1的活性以的活性以及改变细胞周期等方式而促进肺癌细胞的恶性表型。及改变细胞周期等方式而促进肺癌细胞的恶性表型。中文文稿的形成（中文文稿的形成（1 2月）月）导师修改（导师修改（5 6 遍）遍）中文中文 英文英文按预投杂志的稿约整理文章，在线投稿按预投杂志的稿约整理文章，在线投稿论文完成的经过论文完成的经过自己翻译自己翻译自己翻译，找公司润色自己翻译，找公司润色公司翻译并润色公司翻译并润色第一次投往第一次投往Clinical Cancer Rese

9、arch（IF=6.75）Dear Dr. Wang: We regret to inform you that the above-referenced submission is not acceptable for publication in CLINICAL CANCER RESEARCH. Decisions rendered on CLINICAL CANCER RESEARCH submissions are based on the Editor-in-Chiefs, Deputy Editors, or Senior Editors assessment of all of

10、 the information obtained on a manuscript during the submission process. Only those manuscripts that meet stringent requirements of high scientific quality and significance, originality, and priority can be accepted. Based on a preliminary review, it is the opinion of the Editorial Board that your m

11、anuscript does not meet all of these criteria and, therefore, is not likely to be acceptable for publication. 创新性不够创新性不够, , Rejected!分析拒稿原因：分析拒稿原因： 文章的创新点不够突出文章的创新点不够突出 重新组织语言，突出创新重新组织语言，突出创新 Cover Letter书写平淡书写平淡 加入文章主要内容，突出重点加入文章主要内容，突出重点修改前的修改前的Cover Letter：Dear Editors, We submit our manuscript e

12、ntitled “” for possible publication in Clinical Cancer Research. In our study, we found that the mRNA and protein expression of -catenin were increased in lung cancer tissues, and its expression was also correlated with poor prognosis of patients. Moreover, we found that there was no competition bet

13、ween -catenin and p120ctn, although the two proteins can all bind to the same domain of E-cadherin. Furthermore, -catenin promoted malignant phenotype of lung cancer cells through changing small GTPases activity and cell cycle 修改后的修改后的Cover Letter：Dear Editors, We submit our manuscript entitled “” f

14、or possible publication in The Journal of Pathology. -catenin, like p120ctn, is a member of catenin family. However, it is poorly understood about expressive profile and clinical significance of -catenin in many tumors. In our study, we found that the mRNA and protein expression of -catenin were inc

15、reased in lung cancer tissues. In 115 cases of NSCLC specimens, the positive expression of -catenin was closely associated with clinicopathological factors (including pTNM stage, lymph node metastasis, etc), and its expression was also correlated with poor prognosis of patients with follow-up record

16、s. Moreover, we found that there was no competition between -catenin and p120ctn, although the two proteins can all bind to the same domain of E-cadherin in cytoplasm. Furthermore, -catenin promoted malignant phenotype of lung cancer cells through changing small GTPases activity and cell cycle 第二次投往

17、第二次投往The Journal of Pathology（IF=6.47）(Major Revision)Reviewer: 1 1. Include and describe the p120ctn and E-cadherin localization in the 115 cases of NSCLC. It is possible to get a better prognostic factor by using the combined -catenin, p120ctn and E-cadherin.Re: Following the instructions of the r

18、eviewer, we analyzed the expression and localization of -catenin, p120ctn and E-cadherin in serial sections by IHC. The results showed that p120ctn and E-cadherin primarily localized in the cytoplasm of lung cancer tissues (defined as abnormal expression), compared with primarily membrane staining i

19、n normal bronchial epithelial cells. Moreover, statistical analysis demonstrated that abnormal expression rate of p120ctn and E-cadherin were higher in tumors and had a higher degree of malignancy (high-stage, poor differentiation, and nodal metastasis). This data suggest that the abnormal expressio

20、n of p120ctn and E-cadherin was significantly associated with poor prognosis of the NSCLC patients, which was consistent with our previous study (Liu Y, et al.). 2. As mentioned in Lu et al, J Cell Biol 1999, -catenin binds to the JMD of E-cadherin. This JMD is also indicated as a binding place for

21、p120ctn. The authors were surprised that they found no competition between -catenin and p120ctn, but they did not attempt to provide an explanation. They also mention that in the cell lines used, p120ctn, -catenin and E-cadherin interact with each other in the cytoplasm. Please include additional ex

22、periments with (1) cell lines in which both catenins and E-cadherin localize at the PM and (2) some E-cadherin mutants with amino acid substitutions in JMD. With this approach you might be able to find smaller interaction domains -catenin and p120ctn that do not overlap.Re: We thank the reviewer for

23、 giving us such a good idea, Although additional experiments can demonstrate whether there is competition between -catenin and p120ctn for binding to E-cadherin in PM and the specific sites of catenins binding to E-cadherin can be confirmed, they would deviate from our original point in that there i

24、s no competition between -catenin and p120ctn for binding to E-cadherin in cytoplasm. Moreover, it will take a long time to complete these experiments, which already constitutes another complete experimental study; we plan to do that in our future research. 3. The material and method section is not

25、complete. For example, there are no details about the cell cycle, proliferation, and matrigel invasive assay. Without these details it is difficult to follow the results in the figures. Reviewer: 2 1. For Figure 1, while it is convincing that -catenin immunoreactivity is increased in lung cancer, it

26、 is not clear whether E-cadherin and p120ctn are down-regulated in the same tumor cluster. The adjacent sections should be compared to indicate this relationship. This is important because if -catenin interacts with E-cadherin and p120ctn in the cytoplasm, one would expect these proteins express in

27、the same subcellular compartments in human lung cancer tissues.Re: Following the instructions of the reviewer, we detected the expression and localization of -catenin, p120ctn and E-cadherin in serial sections by IHC. The results showed that p120ctn and E-cadherin primarily localized in cytoplasm of

28、 lung cancer tissues (defined as abnormal expression), compared with primarily membrane staining in normal bronchial epithelial cells. Nevertheless, the immunoreactivity of E-cadherin and p120ctn did not decrease significantly in lung cancer tissues. 2. A second anti-catenin antibody should be used

29、to confirm at least some of the tissue IHC and Western blot data. Because -catenin expression in lung cancer is not well-characterized, one should be cautious regarding the authenticity of the antibody against -catenin. Re: In order to examine the authenticity of antibody, another new -catenin antib

30、ody purchased from Abcam Ltd. was used to confirm the results of some of IHC and WB. The results of IHC showed that -catenin immunoreactivity was also increased in lung cancer tissues by the use of this new antibody, just like the other -catenin antibody (Santa Cruz). This new -catenin antibody (Abc

31、am) was consistent with -catenin antibody from Santa Cruz and also showed identical MW protein band at approximate 133kDa in lung cancer cell lines or tissues. 3. For Figure 4, reverse immunoprecipitation using anti-delta-catenin would strengthen the statement that the interactions between delta-catenin or p120ctn and E-cadherin a