Synthesis, Anti-Proliferative Activity Evaluation and 3D-QSAR Study of Naphthoquinone Derivatives as Potential Anti-Colorectal Cancer Agents.

Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase Chain Reaction Method

This study was designed to investigate the antioxidant and antiproliferative activities of water and methanol extracts of endemic plant Gypsophila sphaerocephala subsp cappadocica in conjuction with phenolic profiles. Individual phenolics of the extracts were identified and quantified by RP-HPLC analysis. Antioxidant potentials of the extracts were evaluated by DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS [2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] radical scavenging capacity tests, cupric ion reducing antioxidant capacity (CUPRAC) method and Fe2+ chelating assay. Antiproliferative activities of the extracts were tested against MCF-7 (breast adenocarcinoma), HT-29 (colorectal adenocarcinoma) and HepG2 (hepatocellular carcinoma) cell lines. RP-HPLC analysis showed that methanol extract was richer than water extract in terms of phenolic content. In parallel to the phenolic contents, methanol extract showed higher antioxidant activity than water extract by DPPH, CUPRAC and Fe2+ chelating tests while water extract exhibited higher activity by ABTS method. Moreover, methanol extract displayed 1.8-fold, 4.3-fold and 2.6-fold more antiproliferative activity than water extract against MCF-7 cells, HepG2 cells and HT-29 cells, respectively. However, both extracts were found to show moderate antioxidant and antiproliferative activity compared to positive controls. These results suggest that Gypsophila sphaerocephala may be used as a promising source for food and nutraceutical industries due to its considerable antioxidant and antiproliferative potentials together with its rich phenolic content.

Latest statistics reported by Taiwan’s Health Promotion Administration, malignancy has been the major causes of death in Taiwan for 30 years; among those, colorectal cancer is the first-leading cause of cancer deaths. Meanwhile, colorectal cancer is the 3rd most common cancer worldwide. In contrast to other cancers, development of colorectal cancer is an ordered event of genetic and epigenetic changes that called adenoma-carcinoma sequence. Our laboratory cooperates with Dr. Kuo-Hsiung Lee (Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC, USA) and Dr. Jing-Ping Liou (School of Pharmacy, College of Pharmacy, Taipei Medical University, Taiwan) to evaluate the anticancer effect and the underlying mechanisms of DYZ-2-90 and MPT0G030 in human colorectal cancer, respectively. In situations of aberrant hyperproliferation, DYZ-2-90 targets to the mitotic spindles, the critical structure of cell mitosis, and leads to microtubule depolymerization in HT-29 cells. DYZ-2-90 is a novel ring-opened compound modified from neo-tanshinlactone isolated from Chinese medicinal herb Tanshen. It binds directly to microtubules and rapidly suppresses microtubule polymerization in human colorectal cancer cells. Thus, the alteration of mitotic spindle organization changes the scaffolding properties of microtubules, which induces strong and sustained ERK activation and leads to mitotic arrest. Prolonged ERK activation and cyclin B degradation contributes to maintain the cell mitotic state and protect cells from apoptosis, but meanwhile provides more time to accumulate mitotic stress/cell death signals. Then, the cell apoptosis is triggered by anti-apoptotic protein Mcl-1 degradation and JNK activation, which breach the death threshold. The cell fate of mitotic arrest cells is dictated by two competing networks: one is the cytoprotective ERK pathway, and the other is stressed-related JNK pathway. Whereas, MPT0G030, a HDAC inhibitor, redirects colorectal cancer cells (HT-29 cells) to normal colonic life cycle to undergo cell differentiation and cell apoptosis. MPT0G030 is a potent HDAC inhibitor that showed broad-spectrum cytotoxicity against various human cancer cell lines. It not only effectively inhibits class I HDACs, which are overexpressed in many malignant neoplasms, but also redistributes E-cadherin and activates PKCδ, which is linked to cell apoptosis and differentiation. Further, activation of PKCδ is demonstrated to be modulated through HDAC1. Collectively, MPT0G030-induced PKCδ participates in cell apoptosis and concomitantly promotes differentiation of colon cancer cells through E-cadherin redistribution and changes in cell morphology. In this thesis, our results indicate that both DYZ-2-90, a novel microtubule-destabilizing agent, and MPT0G030, a class I HDAC inhibitor, have great potential as new drug candidates for colorectal cancer therapy.

Purpose : This study aimed to evaluate the possible cytotoxic activity of the essential oil of the Eugenia uniflora leaves on normal and tumoral human cells by colorimetric cell viability assay, based on the use of tetrazolium salt (XTT). Methods : To achieve the cytotoxicity it was used normal human lung fibroblast cell line GMO7492A and the evaluation of antiproliferative activity was performed on three tumor cell lines, as follows: human glioblastoma (MO59J), human cervical adenocarcinoma (HeLa) and human breast adenocarcinoma (MCF-7). For the determination of cytotoxic concentration to the normal line, 12 concentrations were evaluated being from 2.44 to 5000 µg/mL. In the evaluation of antiproliferative activity were tested 8 different concentrations of the extract (3.91 to 500 µg/mL). Results : The results in the normal line GM07492A showed that concentrations higher or equal to 39.1 µg/mL are significantly different of the negative control. In the evaluation of antiproliferative activity on tumor cell lines MO59J, HeLa and MCF-7 was observed that the concentration of 125 µg/mL showed a cytotoxic effect on these lines being significantly different from the negative control. The results from the evaluation of the antiproliferative activity in different tumor cell lines of E. uniflora oil was not selective for the tested cell types. Conclusion : E. uniflora oil did not show cytotoxic activity in concentrations lower than 39.1 µg/mL, however, the values found to IC 50 for tumor cells were superior, concluding that the oil has no selectivity for tumor cells tested.

Thiosemicarbazones and 4-thiazolidinones indole-based derivatives: Synthesis, evaluation of antiproliferative activity, cell death mechanisms and topoisomerase inhibition assay

Published on:June 2022 Journal of Young Pharmacists, 2022; 14(2):198-202 Original Article | doi:10.5530/jyp.2022.14.37 Authors: Sagar Puli, Ramasamy Raveendran* Department of Pharmacology, Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER), Dhanvantari Nagar, Puducherry, INDIA. Abstract: Background: Colorectal cancer (CRC) is the third leading cause of death, probably because of its invasive and metastatic potential. Agents that can mitigate the growth and prevent migration may benefit CRC. Diphyllin and its analogues belong to glycosylated compounds previously reported for their in vitro cytotoxicity against many viruses, candida, and human malignant cells. This study aims to evaluate the anti-proliferative and antimigratory activities of diphyllin on human CRC cells. Methods: The MTT assay was performed to determine the 50% inhibitory concentrations (IC50) in HT-29, SW-480, and HCT-15 CRC cells. The acridine orange and ethidium bromide (AO/EB) dual staining was used to ascertain the type of cell death. HT-29 cells were subjected to short-term 5-fluorouracil+oxaliplatin (5-FU+Ox) to eliminate the most sensitive cells. The remaining surviving cells were further treated with diphyllin alone or combined with 5-FU. The combination effect of diphyllin with 5-FU in 5-FU+Ox surviving HT-29 cells was determined by flow cytometry-based Annexin V apoptosis assay. The wound-healing assay was used to detect the anti-migratory activity of diphyllin on 5-FU+Ox surviving HT-29 CRC cells. Results: The IC50 values of diphyllin were found to be 2.9±0.38, 1.3±0.28, and 3.9±0.65 μg/mL against HT-29, SW-480, and HCT-15, respectively. Microscopic examination manifested apoptotic changes like chromatin condensation, nuclear fragmentation, and formation of apoptotic bodies in diphyllin treated CRC cells. Flow cytometry analysis revealed that diphyllin enhances the apoptosis in 5-FU+Ox surviving HT-29 cells. Wound-healing assay displayed the inhibitory activity of diphyllin in 5-FU+Ox surviving HT-29 cells. Conclusion: Diphyllin shows anti-proliferative activity by inducing apoptosis in HT-29, SW-480, HCT-15 cells and 5-FU+Ox surviving HT-29 cells and impairs their migration. Key words: Anticancer, Apoptosis, Cytotoxicity, Flow cytometry, Woundhealing.

HSV infection has high incidence rate and high prevalence. HSV-1 may cause cold sores, fever blisters, and encephalitis. Moreover, HSV-1 has the ability to cause the latent infection of neurons. Therefore, the QSAR studies of HSV-1 inhibitors and drug design based on QSAR have great importance. This paper focused on the pyrazolo[1,5- a]pyridines, which are not acyclic nucleoside analog. The reported pyrazolopyridines showed potent and selective inhibition activity. In order to find novel compounds with higher activity and to develop better predictive models, comparative molecular field analysis (CoMFA), comparative molecular similarity indices analysis (CoMSIA), and hologram quantitative structure activity relationship (HQSAR) methods were performed on a series of HSV-1 inhibitors. The models built by CoMFA, CoMSIA, and HQSAR were found to be suitable for the compounds investigated with good predictive power (CoMFA q2=0.669, R2=0.953; CoMSIA q2=0.693, R2=0.919; HQSAR q2=0.619, R2=0.921). 3D contour maps obtained from PLS models of CoMFA and CoMSIA, and the atom contribution map from PLS model of HQSAR indicated that small steric volumes and electron-withdrawing groups in the R2, R3 regions would be useful to improve the activity. Results showed that the established models of the present work would be helpful in studying the relationship between the bioactivity and the molecular structures and discovering more potent HSV-1inhibitors.

Background: Colorectal cancer (CRC) is one of the most frequent forms of cancer worldwide. Cells with abnormal growth have the ability to invade or spread to other parts of the body (metastasis). Metastasis is the most common cause of death in CRC patients and activation of epithelial-mesenchymal transition (EMT) triggers metastasis. Thus, it is important to understand the mechanisms of EMT to increase the survival rate of CRC. MicroRNA-17 (miRNA-17) has been proven to be significantly higher in CRC tissues than normal tissues. However, there are few papers about the role of miRNA-17 in CRC metastasis. Therefore, in this study, we evaluated the mechanism underlying miRNA-17-5p related EMT in colon cancer cells. Methods: Expression level of miRNA-17-5p as well as target mRNA were analyzed in HCT-15, HT29, LoVo, SW480 and COLO205 cell lines. In order to evaluate the correlation between miRNA-17-5p and EMT phenomenon, selected cell lines were transiently transfected either miRNA-17-5p mimic or miRNA-17-5p inhibitor and then phenotyping studies were conducted. Moreover, to find out the direct target of miRNA-17-5p, we performed AGO2 immunoprecipitation in each colon cancer cell lines. Results: Real-time PCR revealed that miRNA-17-5p expression reversely correlated with vimentin expression in five colon cancer cell lines. Overexpression of miRNA-17-5p inhibited vimentin expression, cell migration and cell invasion in both LoVo and HT29 cells. On the other hand, inhibition of miRNA-17-5p promoted vimentin expression, cell migration and cell invasion in the same parallel cell lines. Through AGO2 immunoprecipitation analysis, we observed that miRNA-17-5p directly bind to vimentin 3'UTR to influence its stability and expression at the transcriptional level. Conclusion: Our data demonstrated that down-regulated miRNA-17-5p promotes colon cancer cell migration and invasion by regulating vimentin expression. These findings propose that miRNA-17-5p may serve as a potential prognostic biomarker in colorectal cancer. Citation Format: Tea Won Kim, Yeo Song Lee, Hye Kyung Hong, Su Jeong Song, Nak Hyeon Yun, Yong Beom Cho. Downregulation of microRNA-17-5p promotes EMT by targeting vimentin in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 492.

Ziziphora (Cacotti in Persian) belongs to the Lamiaceae family (mint group) and is vastly found in Iran and Asia. This traditional medicinal plant is normally used as analgesic and for treatment of particular gastrointestinal diseases. Since colorectal cancer is one of the most common causes of death in the world and the second leading cause of cancer death among adults, there is a pressing need to inhibit this malignancy by using methods with minimal side effects. One of these methods is the use of natural resources such as medical plants. This study is aimed at investigating the expression of apoptosis-related genes in the adjacent culture of colorectal cancer epithelial cells (HT-29) with Ziziphora essential oil (ZEO). The essential oil was extracted from Ziziphora leaves, and its compounds were determined and then added to the HT-29 culture medium at different concentrations. After 24 hours, the HT-29 cells were harvested from the medium and cytotoxicity was analyzed by MTT assay. After MTT assay and determination of the percentage of apoptosis by flow cytometry, RNA extraction was performed and the expression levels of Bax, Bcl-2, caspase 3 (C3), and caspase 9 (C9) were analyzed using newly designed primers by reverse transcription (RT) qPCR method and GeniX6 software. Also, specific antibodies were used for western blot analyses of those molecules. GC analysis revealed 42 different compounds in the ZEO, including pulegone (26.65%), menthone (5.74%), thymol (5.51%), and menthol (1.02%). MTT assay showed that the concentration of 200 μg/ml of ZEO had the highest HT-29 cell death during 24 hours. After incubation with the concentration of 50 μg/ml of ZEO for 24 and 48 hours, caspase 3 and 9 gene expressions in the treated group increased compared to those in the control group (P < 0.001), while the Bcl-2 expression decreased. The results showed that having anticancer compounds, ZEO can increase C3 and C9 and decrease Bcl-2 expressions, causing apoptosis in HT-29 cells in vitro. This can lead to the use of ZEO as a factor for colorectal cancer treatment.

Colorectal cancer (CRC) is the third leading cause of cancer-related deaths in the United States. Common chemotherapeutic regimens for CRC include a combination of chemotherapeutic agents to produce synergistic drug activity and reduce adverse effects. However, adverse effects due to lack of selective toxicity is still a major problem with many chemotherapeutic agents. Novel drugs that display tumor selective toxicity are desperately needed. Several studies have shown that artemisinin monomers (Clinically used anti-malarial agent) possess antineoplastic activity with minimal toxicity. Interestingly, artemisinin dimers showed more potent antineoplastic activity compared to the monomers. However, few studies have fully characterized the activity of these molecules. In this study, we tested the antitumor activity of five chemically-synthesized dihydroartemisinin (DHA) dimers using the human colon cancer cell lines, HT29 and HCT116 and the non-tumorigenic colon cell line, FHC. Compared to other tested dimers, the DHA oxime dimer, NSC735847 showed pronounced selective toxicity in HT29 and HCT116 cells. Using MTS cell viability assays, NSC735847caused a preferential reduction in the viability of HT29 and HCT116 colon cancer cells compared to the non-tumorigenic FHC cells. In addition, NSC735847 significantly increased caspase 3/7 activity in HT29 and HCT116 cells but not in FHC cells which suggests that this compound causes selective apoptosis in these colon cancer cell lines. The combination of NSC735847 and the topoisomerase I inhibitor, irinotecan (commonly used chemotherapeutic agent for colorectal cancer) showed synergistic antitumor activity in HT29 cells. The combination of the two drugs caused a significant increase in cell death and caspase 3/7 activity which were greater than those caused by the individual drugs. Previous studies using other tumorigenic cell lines, suggested that NSC735847 induces oxidative and endoplasmic reticulum (ER) stress. To gain insight into the potential mechanisms of anti-colorectal cancer activity of NSC735847, we tested if this compound causes ER stress and/ or oxidative stress in HT29 cells and whether ER stress was required for NSC735847-induced apoptotic cell death. NSC735847 increased the expression of ER stress marker C/EBP homologous protein 10 (CHOP10) in HT29 cells and the use of ER stress inhibitor, salubrinal, significantly decreased NSC735847-induced cell death and apoptosis. Additionally, NSC735847 caused oxidative stress in HT29 cells which was inhibited by pretreatment of the cells with the antioxidant, trolox. The crosstalk between oxidative stress and ER stress in NSC735847-induced apoptosis in colon cancer cells is still under investigation. Our results suggest that NSC735847 causes ER stress in HT29 cells which plays a major role in drug-induced cell death and apoptosis in these cells. Citation Format: Ahmed E. Elhassanny, Eman Soliman, Paul McGuire, Mahmoud ElSohly, Waseem Gul, Rukiyah Van Dross. The dihydroartemisinin oxime dimer (NSC735847) displays a selective toxicity in colon cancer cells which is potentially mediated by endoplasmic reticulum stress [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4066. doi:10.1158/1538-7445.AM2017-4066

Background: Colorectal cancer (CRC) is the second-leading death of all cancers in the world. Metastasis is the basis for cause of death in patients with CRC. Currently, combination of anti-VEGF therapy and DNA damaging agents, such as Bevacizumab + FOLFOX, has become the first-line therapy in managing colorectal cancer progression. To mimic the first-line therapy, we have conjugated the angiogenesis inhibitory and the DNA damaging moieties to form a bifunctional hybrid molecule. The purpose of this study is to prove the efficacy and safety of the novel hybrid molecule in treatment of CRC cells in mouse models. Materials and methods: The novel hybrid molecule, designated as BO-2762, was synthesized as previously reported. The anti-proliferative activity was analyzed by PrestoBlue® assay in a variety of CRC cell lines and CRC patient derived organoids (PDO). DNA damage was analyzed by alkaline gel electrophoresis and comet assay. Antiangiogenic property was assessed using the docking model, in vitro tube formation, and in vivo angiogenesis methods. Anti-tumor efficacy of BO-2762 was determined using xenografts of CRC cell lines and PDOs at 20 mg/kg via tail vein injection. For safety profiling and toxicology study, the ICR mice model was adopted for various pathological analyses. Results: The values of anti-proliferative IC50 of BO-2762 against a panel of 11 CRC and 10 PDO cell lines were ranged between 0.5 to 4.5 µM. Subsequently, we demonstrated that BO-2762 induced DNA inter-strand cross-linking (ICL) by alkaline agarose gel electrophoresis. Apparently, ICLs caused significant cell cycle arrest at the S phase, and subsequently triggered apoptosis. The angiogenesis inhibition by BO-2762 was also confirmed by in vitro tube formation assay and Direct In Vivo Angiogenesis Assay. The anti-CRC activity of BO-2762 was assessed using xenografts of CRC cells, derived from metastatic sites (LoVo and SW620 cells) and from primary sites (LS1034 and HT-29 cells). Via tail vein injection of BO-2762 at 20 mg/kg for 10 times, the growth of LoVo, SW620, LS1034, and HT-29 cells was suppressed by 85.8%, 83.0%, 75.4%, and 44.8% respectively. In addition, BO-2762 also significantly suppressed the growth of metastatic PDOs (T53 and T112) by 90%. Biosafety analysis showed promising safety parameters in blood chemistry and major organs. Conclusions: Our present results have shown that conjugated hybrid molecule with DNA cross-linking and anti-angiogenic activity is likely a novel approach to treat patients with CRC. Citation Format: Vaikar Navakanth Rao, Kuo-Chu Lai, Chin-Hung Liu, Shung-Haur Yang, Jeou-Yuan Chang, Tsann-Long Su, Te-Chang Lee. A novel hybrid with anti-angiogenic and DNA crosslinking activities potently suppresses the growth of colorectal cancer cells and patient-derived organoids in mice [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr B010.

To the Editor: The October 2012 issue of the Croatian Medical Journal included an epidemiological study reporting cancer mortality in Turkey (1). Concerning this study, we would like to present epidemiological characteristics of patients who died of cancer in the period 2007-2010 at our center in Istanbul, as additional epidemiological data on cancer mortality in Turkey. We retrospectively analyzed epidemiological characteristics of 255 cancer patients who died between July 2, 2007 and July 2, 2010 at GATA Haydarpasa Training Hospital, Department of Medical Oncology in Istanbul. The focus was on the distribution of the type of cancer depending on sex and age. In addition, a sub-group analysis was performed according to age groups. The most common causes of cancer deaths in men were lung cancer (41.5%), colorectal cancer (16.3%), and stomach cancer (7.5%) and in women breast cancer (27.1%), lung cancer (14.6%), and colorectal cancer (13.6%). When age groups were considered, the most common cause of death among men was Ewing's sarcoma between 20-40 years; lung cancer between 41-60 years and 61-80 years; and colorectal cancer in the group older than 81 years. The most common cause of death among women between 20-40 years, 41-60 years, and 61-80 years was breast cancer, while in the group older than 81 years it was lung cancer. Lung and breast cancers are also most common causes of cancer death in the world (2-5). However, the third most common cause of death among men is prostate cancer, while in our study it was gastric cancer, with prostate cancer ranking fourth. The ranking of causes of death among women in our study was the same as in other studies in the world (2-5). The number of studies on cancer epidemiology in our country is limited. In this regard, studies with larger number of patients that more accurately reflect the general characteristics of the patients in the country are necessary. In order to achieve this, it is necessary to establish a nation-wide and continually updated database.

To explore novel natural product‐based nitrogen‐containing heterocyclic compounds with antiproliferative activity, 20 L‐carvone‐derived pyrimidine‐urea compounds 4a–4t were synthesized through the multi‐step reaction of L‐carvone, and structurally characterized by Fourier transform infrared (FT‐IR), hydrogen‐1 nuclear magnetic resonance (1H‐NMR), Carbon‐13 nuclear magnetic resonance (13C‐NMR), and High‐resolution mass spectrometry (HRMS). Besides, the in vitro antiproliferative activity of the target compounds against HepG2, Hela, and MCF‐7 cells was evaluated by methyl thiazolyl tetrazolium (MTT) assay. According to the results, the target compounds showed certain inhibitory activities against the tested cancer cell lines, and five compounds (4b, 4h, 4k, 4l, and 4t) exhibited better inhibition activities against Hela cells than the positive control (5‐FU). Among them, compound 4b held significant antiproliferative activities against Hela and HepG2 cells, and thus deserved further study as a leading compound of new anticancer drugs. In addition, an effective and reasonable three‐dimensional quantitative structure‐activity relationships (3D‐QSAR) model was built by the Comparative molecular field analysis (CoMFA) method to analyze the relationship between the structures of the target compounds and their antiproliferative activities (expressed as pIC50) against Hela cells, and proven to have good predictive ability. Molecular docking was carried out to study the possible binding modes of compound 4b and Survivin, and it was found that compound 4b could be well embedded into the active site, along with the formation of several hydrogen bonds and hydrophobic interactions.

Colorectal cancer is the third most common cause of cancer-related deaths in the world. Surgical intervention followed by chemotherapy remains the primary approach to treatment since colon cancers remain refractory to most chemotherapeutic agents. Based on that, we established a program to screen natural products for cytotoxic activity, employing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay system utilizing HT-29 human colon cancer cells. During the course of our screening, we found that the methanolic extract of silkworm droppings (SDME) has cytotoxic effects on HT-29 cells. In the present study, we investigated the possible mechanisms by which SDME exerts its antiproliferative activity in HT-29 cells. As expected, SDME inhibited growth of HT-29 cells in a dose-dependent manner as assessed by the MTT reduction assay, the lactate dehydrogenase release assay, and the colony formation assay. We also investigated whether the apoptotic effects induced by SDME involve the caspase pathway using the caspase colorimetric assay. Interestingly, caspase-9 and -3, but not caspase-8, were activated in response to SDME treatment. Taken together, these results clearly indicate that the induction of apoptosis by SDME involves a mitochondrial-mediated pathway and strongly suggest that SDME may potentially be a chemotherapeutic agent for human colon cancer.

Background: Non-cancer causes of death in patients with colorectal cancer (CRC) have not received much attention until now. The purpose of the current study is to investigate the non-cancer causes of death in patients with CRC at different periods of latency.Methods: Eligible patients with CRC were included from the Surveillance, Epidemiology, and End Results (SEER) database, and standardized mortality ratios (SMRs) were calculated using the SEER*Stat software 8.3.8.Results: A total of 475,771 patients with CRC were included, of whom 230,841 patients died during the follow-up period. Within 5 years, CRC was the leading cause of death. Over time, non-cancer causes of death account for an increasing proportion. When followed up for more than 10 years, non-cancer deaths accounted for 71.9% of all deaths worldwide. Cardiovascular diseases were the most common causes of non-cancer deaths, accounting for 15.4% of the total mortality. Patients had a significantly higher risk of death from septicemia within the first year after diagnosis compared with the general population (SMR, 3.39; 95% CI, 3.11–3.69). Within 5–10 years after CRC diagnosis, patients had a significantly higher risk of death from diabetes mellitus (SMR, 1.27; 95% CI, 1.19–1.36). During the course of more than 10 years, patients with CRC had a significantly higher risk of death from atherosclerosis (SMR 1.47; 95% CI, 1.11–1.9).Conclusions: Although CRC has always been the leading cause of death in patients with CRC, non-cancer causes of death should not be ignored. For patients with cancer, we should not only focus on anti-tumor therapies but also pay attention to the occurrence of other risks to prevent and manage them in advance.