Effect of PAK1 gene silencing on proliferation and apoptosis in hepatocellular carcinoma cell lines MHCC97-H and HepG2 and cells in xenograft tumor
Authors: Yu-Ming Jia
Zhi-Lei Zhang
Li Peng
Guang-Chao Liu
Lei Mao
Wu-Han Yang
Chong Zhang
Journal: Gene Therapy (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: PAK1 gene silencing; apoptosis; proliferation; hepatocellular carcinoma; MHCC97-H cells
Affiliations: Department of Hepatobiliary Surgery, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, People's Republic of China., Department of Hepatobiliary Surgery, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, People's Republic of China. .

Abstract:  This study intends to explore the effect of the PAK1 gene silencing on apoptosis and proliferation of hepatocellular carcinoma (HCC) MHCC97-H and HepG2 cells and cells in xenograft tumor. MHCC97-H and HepG2 cells and mice with xenograft tumor in vivo were randomly divided into control, empty vector and PAK1 shRNA groups. Morphology and the expression of green fluorescent protein of MHCC97-H and HepG2 cells and cells in xenograft tumor were observed. MTT assay and flow cytometry were used to detect proliferation, cell cycle and apoptosis of MHCC97-H and HepG2 cells and cells in xenograft tumor. The expressions of PAK1, PCNA, Ki67, Cyclin E, CDK2, p21, p53, Bax and Bcl-2 were measured using the quantitative reverse transcription polymerase chain reaction and western blotting. Compared with the control and empty vector groups, number of adherent cells of MHCC97-H and HepG2 cells and cells in xenograft tumor was reduced, and green fluorescent cells became round and reduced in the PAK1 shRNA group. Cell proliferation, the cells at S phase, the mRNA and protein expressions of PAK1, PCNA, Ki67, Cyclin E, CDK2 and Bcl-2 of MHCC97-H and HepG2 cells and cells in xenograft tumor were decreased, while the cells at G1 phase, apoptosis rate, the mRNA and protein expressions of p21, p53 and Bax of MHCC97-H and HepG2 cells and cells in xenograft tumor were increased in the PAK1 shRNA group. PAK1 gene silencing decreases proliferation of MHCC97-H cells, HepG2 cells and cells in xenograft tumor through the p53/p21 pathway. 

Retracted: No

PubPeer comments:
 This research study was funded by a grant to Butterworth and Reeve, presumably the grant from 2004-2007 related to the u2018Longitudinal study of Dyscalculiau2019. The first article of this 6-yearlong study was published in 2012 (Reeve et al 2012) and the data collection probably started around 2004-2005, however the authors failed to mention explicitly that the same data are used here. The test used by the authors to assess dot enumeration and number comparison is the Dyscalculia Screener (2003) by Brian Butterworth. Should Butterworth have mentioned this? , 
 Please read the Methods section. We explicitly reference that this sample is part of the cohort reported in Reeve et al 2012. Once again, in the Methods section here and in the 2012 paper, it is clear that we did not use the Dyscalculia Screener. In fact, the test is quite different: the Screener uses dot-digit verification, while this study and the 2012 report uses speeded naming of the number of dots. Frankly, we cannot understand why Pseudognaphalium Attenuatum keeps saying that we use the Screener without acknowledging it, when reading the Methods section would make it clear that we did not. So, once again, read the Methods before making this comment. , 
 Thank you for confirming the authors used (very old) data that were collected in 2004 2005 and further analyzed and published in 2018. Your statement contradicts the claim by co-author Robert Reeve who stated in Fritz (International Handbook of Math Learning Difficulties, 2019) p.262 Chapter 16: u201cReeve et al.u2019s (2012) research confirm the validity of Butterworthu2019s (2003) Dyscalculia Screener and show that it is possible to identify children with MLD on the basis of their dot enumeration profiles.u201d ![file] ,