Effects of Echinacoside on Ehrlich Carcinoma in Rats by Targeting Proliferation, Hypoxia and Inflammation.

Background and objectives Ehrlich solid carcinoma (ESC) is a type of tumor originating from a spontaneous mammary adenocarcinoma in mice. It is highly aggressive and fast-growing and can create a solid undifferentiated mass when inserted under the skin. This makes it an ideal model for assessing cancer biology and tumor immunology. Echinacoside is a natural phenylethanoid glycoside with anti-inflammatory, anti-endoplasmic reticulum stress, anti-oxidative stress, and other beneficial properties. This study explored the potential anti-cancer benefits of echinacoside in rats with ESC. The study also analyzed its effects on tumor cell proliferation, differentiation, motility, and inflammation. Methods The study involved injecting rats with tumors in their left hind limb using an intramuscular injection of 2×106 cells. After 14 days, some rats were given a daily intraperitoneal dose of 30 mg/kg echinacoside for three weeks. Muscle samples were then analyzed under an electron microscope. In addition, gene expression and protein levels of various factors such as phosphoinositide 3-kinases (PI3K), mammalian target of rapamycin (mTOR), hypoxia-inducible factor (HIF)-1α, cyclin D1, cyclin-dependent kinase 2 (CDK2), tumor necrosis factor (TNF)-α, and nuclear factor (NF)κB were evaluated in another part of the muscle samples. Results After being treated with echinacoside, the ESC rats experienced a significant increase in their mean survival time from 27 days to 48 days. This treatment also resulted in a decrease in the volume and weight of the tumor. Upon examining the tumor tissue under an electron microscope, signs of damage such as pleomorphic cells, necrosis, nuclear fragmentation, membrane damage with cytoplasmic content spilling, and loss of cellular junction were observed. However, the treatment with echinacoside was effective in improving these effects. Furthermore, the expression of PI3K, mTOR, HIF-1α, cyclin D1, CDK2, TNF-α, and NFκB was significantly reduced due to the echinacoside treatment. Conclusions Our research found that echinacoside has antitumor properties that resulted in a substantial decrease in tumor size and weight, leading to an increase in the average survival time of rats and an improvement in muscle structure. Additionally, echinacoside was shown to ameliorate hypoxia by suppressing HIF-1α, reduce inflammation by decreasing NFκB and TNF-α, decrease proliferation by reducing PI3K, and block cyclin D1 and CDK2 to inhibit differentiation.

Alshehri A ，Albuhayri A ，Alanazi M ，Althubaiti MA ，Aljehani RF ，Alsharif FI ，Alatawi TM ，Albalawi SS ，Khodir AE ，Al-Gayyar MM ... -  《Cureus》

Arctiin Inhibits Inflammation, Fibrosis, and Tumor Cell Migration in Rats With Ehrlich Solid Carcinoma.

ESC or Ehrlich solid carcinoma is a type of tumor originating from a spontaneous mammary adenocarcinoma in mice. It is a highly aggressive and fast-growing carcinoma that can create a solid mass when inserted under the skin. Its solid, undifferentiated form makes it an ideal model for researching cancer biology, tumor immunology, and testing various anti-cancer treatments. Additionally, arctiin has multiple beneficial properties, such as anti-proliferative, anti-oxidative, anti-adipogenic, and anti-bacterial. This study aimed to explore the potential anti-cancer benefits of arctiin in rats with ESC while also analyzing its effects on cell fibrosis markers, tumor cell migration, and inflammasome pathways. Rats were given a tumor in their left hind limb via an intramuscular injection consisting of 2×106 cells. After eight days, some of the rats received a daily oral dose of 30 mg/kg of arctiin for three weeks. Muscle samples were observed under an electron microscope or stained with hematoxylin/eosin. Additionally, gene expression and protein levels of toll-like receptor 4 (TLR4), NLR family pyrin domain containing 3 (NLRP3), signal transducer and activator of transcription 3 (STAT3), transforming growth factor (TGF)-β, endothelial growth factor (VEGF), and cyclin D1 were assessed in another part of the muscle samples. When ESC rats were given arctiin as a treatment, their mean survival time increased and their tumor volume and weight decreased. Additionally, when tumor tissue was examined under an electron microscope, it showed signs of pleomorphic cells, necrosis, nuclear fragmentation, membrane damage with cytoplasmic content spilling, and loss of cellular junction. The stained sections with hematoxylin/eosin showed a dense cellular mass and compressed, degenerated, and atrophied muscle. However, treatment with arctiin improved all these effects. Finally, the expression of TLR4, NLRP3, STAT3, TGF-β, VEGF, and cyclin D1 was significantly reduced with arctiin treatment. Through the use of arctiin, tumor size and weight were effectively reduced, leading to an increase in the average survival time of rats and an improvement in muscle structure. Additional research has shown that arctiin is able to suppress inflammation, fibrosis, and the migration of tumor cells by inhibiting STAT3, TGF-β1, TLR4, NLRP3, VEGF, and cyclin D1.

Alfair BM ，Jabarti AA ，Albalawi SS ，Khodir AE ，Al-Gayyar MM ... -  《Cureus》

Antitumor Activity of Ferulic Acid Against Ehrlich Solid Carcinoma in Rats via Affecting Hypoxia, Oxidative Stress and Cell Proliferation.

Background Ferulic acid is a natural compound commonly found in fruits and vegetables like tomatoes, sweet corn, rice bran, and dong quai. It has various beneficial effects on the body, such as anti-inflammatory, anti-apoptotic, hepatoprotective, cardioprotective, and neuroprotective properties. Aims We conducted a study to investigate the antitumor activity of ferulic acid against Ehrlich solid carcinoma (ESC), specifically by affecting hypoxia-inducible factor (HIF)-1α and its subsequent effects on other factors like nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), cellular Myc (cMyc), cyclin D1, mammalian target of rapamycin (mTOR), and signal transducer and activator of transcription 3 (STAT3). Materials and methods The study involved implanting rats with ESC cells and administering 50 mg/kg of ferulic acid orally daily for eight days. Sections of the muscles with ESC were stained with toluidine blue or immunostained with anti-HIF-1α antibodies. The tumor samples were used to evaluate the expression of HIF-1α, Nrf2, HO-1, cMyc, cyclin D1, mTOR, and STAT3. Results Ferulic acid increased mean survival time, reduced tumor volume and weight, and improved the appearance of the tumor tissue. Furthermore, ferulic acid significantly elevated the expression of Nrf2 and HO-1, while reducing the expression of HIF-1α, Nrf2, HO-1, cMyc, cyclin D1, mTOR, and STAT3. Conclusions Ferulic acid can reduce tumor size and weight while improving the structure of muscle cells, suggesting it may have antineoplastic activity against ESC. Further investigation revealed that ferulic acid downregulates HIF-1α, increasing the expression of antioxidant proteins Nrf2 and HO-1. Additionally, ferulic acid decreases the expression of proliferation markers cMyc and cyclin D1 and downregulates cellular regulators mTOR and STAT3.

Alghamdi MA ，Khalifah TA ，Alhawati HS ，Ruzayq M ，Alrakaf A ，Khodier A ，Al-Gayyar MM ... -  《Cureus》

Ferulic acid inhibits tumor proliferation and attenuates inflammation of hepatic tissues in experimentally induced HCC in rats.

Abdulal ZA ，Altahhan MY ，Qindil AF ，Al-Juhani AM ，Alatawi MA ，Hassan HM ，Al-Gayyar MM ... -  《-》

Antitumor Activity of Ligustilide Against Ehrlich Solid Carcinoma in Rats via Inhibition of Proliferation and Activation of Autophagy.

Background Cancer is the second-leading cause of death worldwide. According to a 2018 WHO report, 9.6 million deaths occurred globally due to cancer. Ehrlich carcinoma is characterized by rapid proliferation and a short survival time. Ligustilide is a phthalide derivative and is one of the main compounds in Danggui essential oil and Rhizoma Chuanxiong. It has many protective effects, such as anticancer, anti-inflammatory, antioxidant, and neuroprotective effects. Aims We conducted this study to investigate the antitumor activity of ligustilide against Ehrlich solid carcinoma (ESC) in rats by affecting beclin 1, mammalian target of rapamycin (mTOR), B-cell lymphoma 2 (BCL2), and 5' AMP-activated protein kinase (AMPK). Materials and methods Twenty rats were intramuscularly implanted in the thigh of the left hind limb with a 200-µL tumor cell suspension in PBS containing 2 × 106 cells. After eight days of inoculation, 10 rats out of the 20 were treated with oral 20 mg/kg ligustilide daily. At the end of the experiment, samples of muscles with ESC were separated. Sections prepared from the muscle samples with ESC were immunohistochemically stained with anti-Ki67 antibodies. Another part of the muscle samples with ESC was used to assess gene expression and protein levels of beclin 1, mTOR, BCL2, and AMPK. Results  Treatment of carcinoma rats with ligustilide elevated the mean survival time and reduced tumor volume and weight. Moreover, examination of tumor tissue stained with hematoxylin/eosin showed an infiltrative, highly cell-dense mass supported by a small to moderate amount of fibrovascular stroma and intersected with multifocal myofibril necrosis. Treatment with ligustilide ameliorated all these effects in the carcinoma group without affecting the control group. Finally, treatment with ligustilide significantly decreased the expression of beclin 1, mTOR, and AMPK associated with elevated expression of BCL2. Conclusions  Our study aimed to explore the potential chemotherapeutic activity of ligustilide against ESC. We found that ligustilide effectively reduced tumor size and weight, indicating its antineoplastic activity against ESC. We further investigated that ligustilide inhibits cell proliferation by suppressing Ki67 and mTOR and activates autophagy through beclin 1 activation. Moreover, ligustilide inhibits apoptosis by upregulating BCL2. Finally, ligustilide reduced the expression of AMPK, preventing its ability to promote tumor cell growth.

Alshehri AF ，Khodier AE ，Al-Gayyar MM 《Cureus》