Hesperetin induces the apoptosis of hepatocellular carcinoma cells via mitochondrial pathway mediated by the increased intracellular reactive oxygen species, ATP and calcium
Authors: Jixiang Zhang
Dandan Wu
Jia Song
Weiguo Dong
Jing Wang
Journal: Medical Oncology (2015)
Discipline: CLINICAL MEDICINE
Keywords: Hesperetin; hepatocellular carcinoma; apoptosis; proliferation; ROS
Affiliations: Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei Province, China.

Abstract:  Hesperetin, a flavonoid from citrus fruits, has been proved to possess biological activity on various types of human cancers. However, few related studies on hepatocellular carcinoma are available. In this study, we aimed to investigate the effect of hesperetin on hepatocellular carcinoma cells in vitro and in vivo and clarify its potentially specific mechanism. Compared with the control group, the proliferations of hepatocellular carcinoma cells in hesperetin groups were significantly inhibited (P &lt; 0.05), and a dose- and time-dependent inhibition of cell viability was observed. When pretreated with H2O2 (1 mM) or N-acetyl-L-cysteine (5 mM), the inhibition of cell viability by hesperetin was enhanced or reduced, respectively (P &lt; 0.05). Similarly, the levels of intracellular ROS, ATP and Ca(2+) changed in different groups (P &lt; 0.05). The results of Hoechst 33258 staining showed that the percentages of apoptotic cells in hesperetin groups are remarkably higher than that in control group (P &lt; 0.05). And the results of Western blot showed that hesperetin caused an increase in the levels of cytosolic AIF, cytosolic Apaf-1, cytosolic Cyt C, caspase-3, caspase-9 and Bax and a decrease in that of Bcl-2, mitochondrial AIF, mitochondrial Apaf-1 and mitochondrial Cyt C (P &lt; 0.05). Meanwhile, hesperetin significantly inhibited the growth of xenograft tumors. Our study suggests that hesperetin could inhibit the proliferation and induce the apoptosis of hepatocellular carcinoma via triggering the activation of the mitochondrial pathway by increasing the levels of intracellular ROS, ATP and Ca(2+). 

Retracted: No

PubPeer comments:
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 More in Fig 7C ![file] , 
 Well I do say, peeps, you jolly well could be onto something here. I donu2019t mean to spoil your fun bigging up the figure details but might I suggest that you also download the pdf for the original submission as presented at JBCu2019s web site? Because, if you do that, teasing apart the lane slices in the Pdf editor of your choice is rather easy peasy and these figure parts are just so explodable... ![image] , 
 #9 Indeed, examining the original submission reveals a multiply-spliced panel in Fig 4D that would otherwise be hard to detect. Adjusting the image levels further reveals some repetitive elements: ![file] , 
 Also, some of these figures have issues other than extensive splicing. In particular, several contain regions with apparent smudging and dodging features. See Fig 5C: ![file] Also Fig 6A: ![file] , 
 #10 As you say, Fig. 4D WB alphaMyc PAN2 is a confection of lane splicing, even if the pasted-in parts donu2019t line up very well. And is that a painted-on background behind them? Of course, the duplication of lanes 4 and 5 was already noted in comment #5 a mere three years ago by Peer 1, not that anything has happened since. Which leads one to wonder whether the current and very excellent JBC Data Integrity Manager has now been contacted, given that there are these new findings and still more to document? As it happens, the as yet uncommented Fig. 4C lanes are all individually spliced into a pre-prepared box too. I wonder if this practice is stated in the Materials and Methods? Here are the parts of these Burnham Institute Western Blots, which get selected when the cursor is moved diagonally across the figure in the original submission ![image] , 
 #12 Excellent observations, Amazonicum. I also like how evident it all looks under your orange-tinted scope. Do you have any particular PDF editor that you d like to share with the audience? , 
 #13 I assume there will be many image editors out there that can extract the objects from a pdf file. These particular orange-tinted selections were done with Preview, the Macu2019s standard pdf editor. I also have an older version of Acrobat Pro which I also use from time to time. I do not plan to rent the more recent offerings from Adobe, so if these two collectively fail me at some point in the future, I would then have to evaluate alternatives. , 
 #12 * Which leads one to wonder whether the current and very excellent JBC Data Integrity Manager has now been contacted... * Yes, it would be good if PubPeer provided an option to alert the journal. Separately, in the  Authors emails section at top right, the e-mail for the corresponding author is obsolete, and has been so for roughly 6 years . It is unfortunate that there is no mechanism to provide an updated address. This issue has been raised in the Bug Report thread: https: pubpeer.com publications bugreport#52 https: pubpeer.com publications bugreport#55 ,