Hwang-Heuk-San induces apoptosis in HCT116 human colorectal cancer cells through the ROS-mediated activation of caspases and the inactivation of the PI3K/Akt signaling pathway
Authors: Su-Hyun Hong
Sang Hoon Hong
Cheol Park
Yung Hyun Choi
Taeg Kyu Kwon
Sun-Hee Leem
Jin Won Hyun
Sung Hyun Choi
Moon Hee Lee
Young-Sam Keum
Gi-Young Kim
Journal: Oncology Reports (2016)
Discipline: CLINICAL MEDICINE
Keywords: Hwang-Heuk-San; polyherbal formulation; gastrointestinal malignancy; anticancer effects; HCT116 cells
Affiliations: Department of Biochemistry, Dongeui University College of Korean Medicine, Busan¬†614-052, Republic of Korea., Department of Molecular Biology, Dongeui University, Busan¬†614-714, Republic of Korea., Department of Marine Life Sciences, Jeju National University, Jeju¬†690-756, Republic of Korea., Department of Biological Science, Dong-A University, Busan¬†604‚Ä?14, Republic of Korea., Department of Safety and System Management, Korea Lift College, Geochang¬†670-802, Republic of Korea., College of Pharmacy, Dongguk University, Goyang¬†410-773, Republic of Korea., Department of Biochemistry, School of Medicine, Jeju National University, Jeju¬†609-756, Republic of Korea., Department of Immunology, School of Medicine, Keimyung University, Daegu¬†704-701, Republic of Korea., Department of Internal Medicine, Dongeui University College of Korean Medicine, Busan¬†614-052, Republic of Korea.

Abstract:  Hwang-Heuk-San (HHS) is a polyherbal formulation that has been used in traditional Korean medicine for hundreds of years to treat gastrointestinal malignancy. However, to date, the mechanisms responsible for the anticancer effects remain unclear. In the present study, we investigated the anticancer effects of HHS using HCT116 human colorectal cancer (CRC) cells. Our results showed that HHS treatment significantly reduced cell survival and increased apoptotic cell death in a concentration-dependent manner. The treatment of HCT116 cells with HHS also significantly elevated the accumulation of reactive oxygen species (ROS), which was followed by the attenuation of the mitochondrial membrane potential through the upregulation of Bax and the downregulation of Bcl-2, which was accompanied by the release of cytochrome c to the cytosol. In addition, HHS treatment caused the truncation of Bid and activated the caspases (caspase-8, -9 and -3), which was associated with the induction of the Fas ligand, the death receptors (DRs), DR4 and DR5, downregulation of the inhibitors of protein expression in the apoptosis protein family, and the degradation of poly(ADP-ribose)-polymerase. However, a pan-caspase inhibitor reversed the HHS-induced apoptosis and growth suppression, indicating that HHS induces apoptosis though a caspase-dependent intrinsic and extrinsic apoptotic pathway in HCT116 cells. Moreover, HHS treatment inhibited the activation of phosphatidylinositol-3-kinase (PI3K)/Akt signaling, and a pharmacological inhibitor of PI3K significantly potentiated the apoptotic effects of HHS when employed in combination in HCT116 cells. Furthermore, the blocking of ROS generation by antioxidant N-acetyl cysteine attenuated the HHS-induced release of cytochrome c, caspase activation and PI3K/Akt inactivation, thereby preventing HHS-induced apoptosis and reduction in cell viability. These findings suggest that HHS-induced ROS generation is required for caspase-dependent apoptotic cell death involving inhibition of the PI3K/Akt signaling pathway in HCT116 cells. Overall, our findings suggest that HHS may be an effective treatment for CRC cancer, and further studies are required to identify the active compounds in HHS. 

Retracted: No

PubPeer comments:
 We have contacted the first author. The first author has indicated that he is now out of raw data, so this makes it impossible to show the process from raw data to intermediate samples. We have contacted and communicated with multiple researchers from multiple universities. None of us was able to reproduce the results. In addition, there are many writing and logical errors in this ICWS paper. These make it impossible for us to understand the details of the paper. We hope you will initiate an investigation and pay enough attention to it! Best, Wei Zhang The following is my conversation with the author: From: Lingzhi Wang lingzhiwang2025@u.northwestern.edu Sent: Friday, 30 April 2021 20:39 To: Zhang Wei WeiZhang110@hotmail.com Subject: Re: Questions about Root-Cause Metric Location for Microservice Systems via Log Anomaly Detection I think with the proper raw data, the final result is reproducible. But I can t show you the first part of work (Log Anomaly Detection). Because I don t have the data and codes (which is not based on my original work). You can also list the  errors you said in my paper if it is convenient for you. And I will appreciate it. Lingzhi On Fri, Apr 30, 2021 at 8:33 PM Zhang Wei WeiZhang110@hotmail.com wrote: Thank you for your reply. I think the  prove is  reproduce . In other words, due to the lack of original files, can t you show me the reproduced process from original files to intermediate data? I want to reproduce the results of your paper. I want to reproduce the whole process. According to you, is it not possible to reproduce the intermediate process of getting the results in your article due to the lack of your original file? I don t know what is wrong with my mailbox. I am sorry for this. From: Lingzhi Wang lingzhiwang2025@u.northwestern.edu Sent: Friday, 30 April 2021 20:22 To: Zhang Wei WeiZhang110@hotmail.com Subject: Re: Questions about Root-Cause Metric Location for Microservice Systems via Log Anomaly Detection What do you mean by  prove ? I cannot show you how I got the intermediary results because I don t have the original log text files now (which may be up to 200+GB). But I can show you some details about how we get it. The logs we have are collected among an experimental micro-services system, which is based on this open-source project https: github.com FudanSELab train-ticket. The process from the original logs to the Log Anomaly Score is based on this paper  Deeplog: Anomaly detection and diagnosis from system logs through deep learning , which is not my work. The KPI monitoring data we have is collected by the tools istio and Prometheus . Did you use the same tools to get your Anomaly Score? I believe this may lead to some problems with your performance. By the way, could you please check what s wrong with your email account? It keeps sending me many emails with the same content. Best regards, Lingzhi On Fri, Apr 30, 2021 at 8:00 PM Zhang Wei WeiZhang110@hotmail.com wrote: why do not reply me? I want to know, can you not prove the process from raw data to intermediate results now? Zhang Wei WeiZhang110@hotmail.com Friday, 30 April 2021 19:39 uff1a Thank you for your reply! As you said, you have no original data. You have no way to prove that it is reasonable to follow your steps from the original data to the intermediate data. This has brought great difficulties to our understanding and further application. In fact, we spent a lot of time to reproduce, but there is a big gap between your results. We donu2019t know why. Due to the companyu2019s agreement, I cannot give you our data. From: Lingzhi Wang lingzhiwang2025@u.northwestern.edu Sent: Friday, 30 April 2021 19:32 To: Zhang Wei WeiZhang110@hotmail.com Subject: Re: Questions about Root-Cause Metric Location for Microservice Systems via Log Anomaly Detection I can reproduce my result based on the intermediary data and results I got, including Log Anomaly Score and KPI monitoring indicators. But now I don t have the original log text files. Lingzhi Zhang Wei WeiZhang110@hotmail.com Friday, 30 April 2021 19:28 uff1a Thank for your reply! In other words, can t you reproduce your results because you lack the original data? From: Lingzhi Wang lingzhiwang2025@u.northwestern.edu Sent: Friday, 30 April 2021 19:18 To: Zhang Wei WeiZhang110@hotmail.com Subject: Re: Questions about Root-Cause Metric Location for Microservice Systems via Log Anomaly Detection I will try my best to give you the data I used(some intermediary data). However, as the raw data is collected by the experimental system in a company, you may need to construct this system by yourself to get the raw data as well as perform log analysis on it. I will share some technical details about the system later. As far as I know, you may need some tools like Istio and Prometheus to collect the raw data. Best regards, Lingzhi Zhang Wei WeiZhang110@hotmail.com Friday, 30 April 2021 19:09uff1a Thank for your reply! I also hope that you can send me your used dataset in this paper. Can you give me your dataset? I would appreciate it if it would be convenient for you to give it to me. Best, Wei Zhang From: Lingzhi Wang lingzhiwang2025@u.northwestern.edu Sent: Friday, 30 April 2021 18:18 To: Zhang Wei WeiZhang110@hotmail.com Subject: Re: Questions about Root-Cause Metric Location for Microservice Systems via Log Anomaly Detection That s Okay. I will send you the code when I go back home tonight. Best regards, Lingzhi Zhang Wei WeiZhang110@hotmail.com Friday, 30 April 2021 18:10uff1a Dear Dr. Wang , Hello. I am Wei Zhang from Xidian University, China. I read your paper titled  Root-Cause Metric Location for Microservice Systems via Log Anomaly Detection . I try to reproduce your results show in this paper for a long time but no progress. Besides, your code is not available. This makes it very difficult for me to verify your paper. I have contacted and communicated with multiple researchers from multiple universities. None of us was able to reproduce your results. In addition, there are many writing and logical errors in your paper. These make it impossible for us to understand the details of your paper. Therefore, if it is convenient, can you give me the your code and datasets about your paper? I promise to use it only for academic research. WE look forward to hearing from you soon! Best, Wei Zhang ,