Inhibition of Drp1 orchestrates the responsiveness of breast cancer cells to paclitaxel but insignificantly relieves paclitaxel-related ovarian damage in mice.

Chemoresistance and chemotherapy-related ovarian damage are well-reported in breast cancer (BC) young patients. Herein, the inhibition of the mitochondrial fission was invested to explore its chemosensitizing role in Paclitaxel (PTX)-resistant cells, and its ability to restore the ovarian integrity in mice receiving PTX or cisplatin chemotherapy. To establish these aims, PTX-resistance was generated in BC cells, which were treated with PTX in combination with Drp1 deficiency, via mdivi-1, or Drp1-specific siRNA transfection. Furthermore, the alterations in the ovarian structure and the endocrine-related hormones were explored in mice receiving repetitive doses of PTX or cisplatin. We found that combining PTX with mdivi-1 improved cell responsiveness to PTX, induced apoptosis- and autophagy-mediated cell death, and relieved cellular oxidative stress. Additionally, the expression of PCNA1 and cyclin B1 genes were downregulated, meanwhile, p53, p21, and mitochondrial fusion proteins (Mfu1&Mfu2) were increased. The in vivo investigations in mice demonstrated that PTX induced gonadotoxic damage similar to cisplatin, whereas dual treatment of mice with PTX+ mdivi-1 failed to restore their normal follicular count and the circulating levels of E2 and AMH hormones. These results suggested that combining Drp1 inhibition with PTX resensitized breast cancer cells to PTX but failed to offer enough protection against chemotherapy-related gonadotoxicity.

Cytotoxic Potential of Novel Quinoline Derivative: 11-(1,4-Bisaminopropylpiperazinyl)5-methyl-5H-indolo[2,3-b]quinoline against Different Cancer Cell Lines via Activation and Deactivation of the Expression of Some Proteins.

The current study evaluated the cytotoxic activity of 11-(1,4-bisaminopropylpiperazinyl)5-methyl-5H-indolo[2,3-b]quinoline (BAPPN), a novel derivative of 5-methyl-5H-indolo[2,3-b]quinoline, against hepatocellular carcinoma (HepG2), colon carcinoma (HCT-116), breast (MCF-7), and lung (A549) cancer cell lines and the possible molecular mechanism through which it exerts its cytotoxic activity. BAPPN was synthesized and characterized with FT-IR and NMR spectroscopy. The binding affinity scores of BAPPN for caspase-3 PDB: 7JL7 was -7.836, with an RMSD of 1.483° A. In silico screening of ADME properties indicated that BAPPN showed promising oral bioavailability records in addition to their high gastrointestinal absorption and blood-brain barrier penetrability. BAPPN induced cytotoxicity, with IC50 values of 3.3, 23, 3.1, and 9.96 µg/mL against cancer cells HepG2, HCT-116, MCF-7, and A549, respectively. In addition, it induced cell injury and morphological changes in ultracellular structure, including cellular delayed activity, vanishing of membrane blebbing, microvilli, cytoplasmic condensation, and shrunken nucleus with more condensed chromatin autophagosomes. Furthermore, BAPPN significantly increased the protein expression of caspase-3 and tumor suppressor protein (P53). However, it significantly reduced the secretion of vascular endothelial growth factor (VEGF) protein into the medium and decreased the protein expression of proliferation cellular nuclear antigen (PCNA) and Ki67 in HepG2, HCT-116, MCF-7, and A549 cells. This study indicates that BAPPN has cytotoxic action against liver, colon, breast, and lung cancer cell lines via the up-regulation of apoptotic proteins, caspase-3 and P53, and the downregulation of proliferative proteins, VEGF, PCNA, and Ki67.

Design and cytotoxic evaluation via apoptotic and antiproliferative activity for novel 11(4-aminophenylamino)neocryptolepine on hepatocellular and colorectal cancer cells.

The current study evaluated the cytotoxic activity of 11(4-Aminophenylamino)neocryptolepine (APAN), a novel derivative of neocryptolepine, on hepatocellular (HepG2) and colon (HCT-116) carcinoma cell lines as well as, the possible molecular mechanism through which it exerts its cytotoxic activity. The APAN was synthesized and characterized based on their spectral analyses. Scanning for anticancer target of APAN by Swiss software indicated that APAN had highest affinity for protein tyrosine kinase 6 enzyme. Furthermore, Super pred software indicated that APAN can be indicated in hepatic and colorectal cells with 92%. Molecular docking studies indicated that the binding affinity scores of APAN for protein PDB code: 6CZ4 of tyrosine kinase 6 recorded of - 6.6084 and RMSD value of 0.8891°A, while that for protein PDB: 7JL7 of caspase 3 was - 6.1712 and RMSD of 0.8490°A. Treatment of HepG2 and HCT-116 cells with APAN induced cytotoxicity with IC50 of 2.6 and 1.82 µg/mL respectively. In addition, it induced injury and serious morphological changes in cells including, disappearance of microvilli, membrane blebbing, cytoplasmic condensation, and shrunken nucleus with more condensed chromatin. Moreover, APAN significantly increased protein expression of annexin V (apoptotic marker). Furthermore, APAN significantly increased protein expression of caspase 3 and P53. However, it significantly reduced secretion of VEGF protein into the medium and decreased protein expression of PCNA and Ki67 in HepG2 and HCT-116 cells. This study indicated that APAN had cytotoxic activity against HepG2 and HCT-116 cells via increasing the expression of apoptotic proteins and reducing the expression of proliferative proteins.

Necroptosis modulation by cisplatin and sunitinib in hepatocellular carcinoma cell line.

Aim Hepatocellular carcinoma (HCC) is one of the leading causes of cancer death worldwide. Systemic chemotherapy such as cisplatin and multi-targeted receptor tyrosine kinase inhibitors, including sunitinib, has marginal activity and frequent toxicity. Recently, necroptosis has been investigated as a potential target in treating cancer. Our aim is to evaluate the influence of cisplatin-sunitinib combination on HepG2 cells regarding their cytotoxicity and implicated intracellular pathways. MATERIALS AND METHODS: The half-maximal inhibitory concentration (IC50) values of cisplatin, sunitinib, and their combination were determined by Sulforhodamine-B assay. Bcl-2 and Bax protein levels were assayed using western blot. ELISA technique was used to measure pRIPK3/RIPK3, pERK/ERK, caspase-9, caspase-8, malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GPx). KEY FINDINGS: Cisplatin-sunitinib combination exhibited a superior cytotoxic effect on HepG2 cells. Low concentrations of 4 µg/ml cisplatin and 2.8 µg/ml sunitinib showed significant Bcl-2 down-regulation and Bax up-regulation. The combined treatment also lowered pRIPK3/RIPK3 by 74% (p < 0.05) compared to the control. Significant increase in pERK/ERK by 3.9 folds over the normal control was also demonstrated. Moreover, combined treatment produced a significant 4 and 4.6 folds increase in caspase-9 and -8 levels. An increase in MDA level by 1.3 folds, a decrease in the intracellular GSH level by 63%, and an increase in GPx level by 1.17 folds were demonstrated. SIGNIFICANCE: Sunitinib modulated cisplatin effect on cytotoxicity, oxidative stress, apoptosis, necroptosis and MAPK pathways. Sunitinib enhanced cisplatin-induced apoptosis and increased oxidative stress, but decreased necroptosis. Combined cisplatin and sunitinib might be promising for treating advanced HCC.

Aldo-keto reductase and sorbitol dehydrogenase enzymes in Egyptian diabetic patients with and without proliferative diabetic retinopathy.

BACKGROUND: Activation of the polyol pathway due to increased aldo-keto reductase (AKR) activity has been implicated in the development of diabetic complications, including proliferative diabetic retinopathy (PDR); however, the relationship between hyperglycaemia-induced activation of the polyol pathway in the retina and PDR is still uncertain. METHODS: This study was conducted on 73 individuals, who were categorised into three groups: healthy individuals as normal control (15 age-matched subjects), diabetic patients treated with oral hypoglycaemic drugs (OHD, 34 patients), six of whom (17.7 per cent) were diagnosed with PDR and the rest were diagnosed with non-proliferative diabetic retinopathy (NPDR) and diabetic patients treated with insulin (INS, 24 patients), 12 of whom (50 per cent) were diagnosed with PDR and the rest had NPDR. RESULTS: The AKR level in diabetic subjects showed a significant increase compared with the normal controls. Interestingly, AKR levels were significantly increased in the INS compared with the OHD group. Also the AKR level was significantly increased in the patients with proliferative compared with the non-proliferative retinopathy in both the insulin and oral diabetic groups. The sorbitol dehydrogenase (SDH) level in diabetic patients showed a significant decrease compared with the normal control level. Interestingly, the SDH level was significantly decreased in the INS compared with the OHD group. Also, the SDH level was significantly decreased in patients with proliferative compared with non-proliferative retinopathy in both INS and OHD groups. The HbA(1c) level in both INS and OHD subjects showed a significant increase compared with normal controls. In addition, the triglyceride level in insulin proliferative retinopathy showed a significant increase compared with other groups. CONCLUSIONS: The AKR level was significantly increased in patients with proliferative compared with non-proliferative retinopathy in both insulin and oral diabetic groups. The SDH level was significantly decreased in patients with proliferative compared with non-proliferative retinopathy in both insulin and oral diabetic groups. Both AKR and SDH could be used as indicators for diabetic control.