Objective： To investigate the mechanism of DEHP-induced damage on mouse spermatogonial cell GC-1 spg function. Methods： GC1 spg were treated with various concentrations of DEHP (0, 30, 60, 90, 120 μmol/L). The effects of DEHP on the proliferation and migration of GC-1 spg were observed by CCK8 test, colony formation test and Transwell assay, and the relative contents of iron ion (Fe3+) and malondialdehyde (MDA) in GC-1 spg cells were determined by chemical spectrophotometry. The expression of cystine glutamate reverse transporter (xCT) and glutathione peroxidase 4 (GPX4) in GC1 spg was detected by Western blotting assay. The expression of intracellular reactive oxygen species （ROS） and the changes of mitochondrial membrane potential were detected by cellular immunofluorescence. The proliferation ability of GC-1 spg was detected by CCK8 assay after ferroptosis inhibitor Ferrostatin1 (1 μmol/L) was cotreated with DEHP (90 μmol/L) 24 h. Results： Compared with the control group (0 μmol/L), the proliferation, cloning and migration ability of cells in 30, 60, 90, 120 μmol/L DEHP groups were significantly decreased, while the contents of MDA,ROS and Fe3+ (90, 120 μmol/L DEHP group) were significantly increased. The mitochondrial membrane potential (60, 90, 120 μmol/L DEHP group) and the expression of GPX4 (60, 90, 120 μmol/L DEHP group), xCT protein were significantly decreased. The ability of cell proliferation after co-treatment of DEHP and Ferrostatin-1 was significantly higher than that of DEHP alone. Conclusion： DEHP can inhibit proliferation and migration capacity of GC-1 spg and reduce the expression of GPX4 and xCT proteins in GC-1 spg, possibly causing ferroptosis.

 ［Abstract］Objective： To explore the role and mechanism of Connexin43 (Cx43), a gap junctional protein of astrocytes (ASC), in regulating apoptosis and ferroptosis. Methods： ASC was treated with 25 μmol/L hemin to establish a cell model that mimics apoptosis and ferroptosis after intracerebral hemorrhage. Lentivirus was used to knock down the expression of Cx43 in ASC, and the sensitivity of cells to hemininduced apoptosis and ferroptosis was evaluated. Through gene set enrichment analysis (GSEA), the possible mechanism of Cx43 involved in the regulation of apoptosis and ferroptosis was predicted. The signaling pathway predicted by the bioinformatics analysis was verified by immunofluorescence staining of Yes associated protein (YAP) and intervention with the YAP inhibitor Verteporfin (Vp). Results： Compared with the control group, hemin treatment caused a significant increase in the proportion of apoptosis and ferroptosis (P<0.05). The Cx43 knockdown group (Cx43KD) had a significantly lower percentage of apoptosis (P<0.05), and higher percentage of ferroptosis (P<0.05). Bioinformatics analysis showed that Cx43 was significantly related to Hippo signaling pathway, iron binding gene set, lipid peroxidation pathway, hypoxic stress pathways. Further transcription factor regulatory network analysis found that TEAD3 in the TEAD transcription factor family may be involved. Immunofluorescence staining showed that YAP was translocated to the nuclei in the Cx43KD group. Blocking the combination of YAP and TEAD with Vp, the apoptosis resistance and ferroptosis sensitization induced by Cx43 knockdown were partially reversed. Conclusion： Cx43 knockdown exerts ASC with resistance to hemin-induced apoptosis and the sensitivity to hemin-induced ferroptosis by inhibiting the Hippo pathway.

Objective: To investigate the effect of gossypol acetate （GAA） on ferroptosis in human glioma cells. Methods: Human brain glioma U87MG, LN229 and U251MG cells in logarithmic growth phase were selected and divided into two groups, treated with 0 or 10 μmol/L GAA, respectively, CCK-8 and clony formation assays were used to detect cell proliferation abilities. QRT-PCR and Western blotting were used to detect the expression of glutathione peroxidase 4（GPX4） in mRNA and protein levels, respectively. Also, the contents of reduced glutathione （GSH） and malondialdehyde （MDA） were detected by chemiluminescence. In addition, three kinds of human glioma cells were co-cultured with 0 or 10 μmol/L GAA combined with 0, 1 μmol/L Ferrostatin-1, respectively, and the cell proliferation rate was detected by CCK-8 method. Results: Compared with 0 μmol/L GAA group, the relative proliferation rate and clonal formation ability of U87MG, LN229 and U251MG cells in GAA 10 μmol/L group were significantly decreased （all P<0.05）, the expression levels of GPX4 mRNA and protein in LN229 and U251MG cells decreased significantly （all P<0.05）, the content of GSH decreased significantly and the content of lipid peroxidation product, malondialdehyde increased remarkably（P<0.05）, while the above indexes in U87MG cells did not change greatly （P>0.05）. Compared with 10 μmol/L GAA+0 μmol/L Ferrostain-1 group, the relative proliferation rate of three brain glioma cells in 10 μmol/L GAA+1 μmol/L Ferrostatin-1 group was significantly increased（all P<0.05）. Conclusion: Gossypol acetate promotes ferroptosis in glioma cells, and LN229 and U251MG cells are more sensitive than U87MG cells.
［Key words］glioma; gossypol acetate; ferroptosis;Ferrostatin-1; glutathione peroxidase 4（GPX4）

铁死亡（ferroptosis）是一种新近定义的细胞死亡形式，与细胞凋亡、自噬以及其他形式的细胞死亡不同，其涉及铁和活性氧。目前研究表明，铁死亡与多种疾病的病理细胞死亡相关，如阿尔茨海默病、脑出血、创伤性脑损伤和缺血再灌注损伤等。此外，在多种肿瘤细胞中也发现铁死亡，其可能具有抑癌作用，并可用于治疗癌症。本文综述细胞铁死亡的可能机制，并回顾铁死亡在肝癌、卵巢癌、头颈部肿瘤以及其他肿瘤中的研究进展。

铁死亡(ferroptosis)是近年来新发现的一种细胞死亡方式，它与细胞内的氧化还原稳态失衡相关。与凋亡、坏死等传统的死亡方式不同，细胞内游离铁增多和脂质过氧化是铁死亡独特的代谢特征。大量研究表明，相比于正常细胞，肿瘤细胞需要更多的铁来维持生长，该特征导致肿瘤细胞对于铁死亡更加敏感。自发现铁死亡以来，学者们陆续提出利用铁死亡的方式杀伤肿瘤的治疗策略。本文针对铁死亡发生过程中肿瘤细胞内铁、脂质和氨基酸代谢的主要调控机制，以及其在胰腺导管细胞癌、肝细胞癌、胶质瘤和血液系统肿瘤中的研究进展进行归纳总结。

Objective: To investigate whether metformin can reverse hypoxiainduced resistance to ferroptosis in pancreatic cancer cells and to further clarify its mechanism. Methods: Human pancreatic cancer cells Patu8988 were cultured under normoxia and hypoxia conditions for different time (0, 12, 24 or 48 h). CCK8 assay was used to detect the cell viability and Western blotting was used to detect the protein expression of hypoxia-inducible factor 1α (HIF1α); CCK8 assay was used to detect the cell viability of Patu8988 cells treated with different concentrations of ferroptosis inducer Erastin in normoxia and hypoxia conditions; the hypoxiacultured Patu8988 cells were divided into the following four treatment groups: control group, metformin group, Erastin group, metformin+Erastin group. CCK8 assay was used to detect the cell viability, Western blotting was used to detect the protein expression level of glutathione peroxidase 4 (GPX4). Patu8988 cells in hypoxia culture were treated with different concentrations of metformin, and Western blotting was used to detect the protein expression levels of AMPactivated protein kinase (AMPK), mammalian target of rapamycin (mTOR), HIF-1α and other proteins. Results: Compared with the normoxia group, hypoxia inhibited the cell viability in a timedependence manner, and the difference between two groups was the most significant at 48 h (P<0.05). When the concentration of Erastin was 10 μmol/L, the cell viability of the hypoxia group was significantly lower than that of the normoxia group (P<0.05), and 10 μmol/L was the best experimental concentration. Compared with the group treated alone in hypoxia condition, the combined treatment could significantly inhibit the cell viability and reduce the protein expression level of GPX4 (both P<0.05). By activating AMPK, inhibiting mTOR phosphorylation, metformin negatively regulated HIF-1α protein expression level (both P<0.05). Conclusion: Metformin could reduce the accumulation of HIF-1α through the AMPK-mTOR axis, thus reversing hypoxia-induced resistance to ferroptosis in pancreatic cancer cells.

Objective: To investigate the effect of heat shock protein family A member 5(HSPA5) on the chemosensitivity of artesunate in human hepatic carcinoma SMMC7721 cells. Methods: SMMC7721 cells were treated with different concentrations of artesunate, CCK8 assay was used to detect the cell viability, and to screen the best conecntration. SMMC-7721 cells were divided into control group, artesunate group, artesunate+deferaxamine group. CCK8 assay was used to test the cell viability. Flow cytometry was used to detect the level of lipid reactive oxygen species(ROS). The malondialdehyde detection kit was used to detect the level of malondialdehyde. qRT-PCR and Western blotting were used to detect the mRNA and protein expression level of HSPA5 in SMMC-7721 cells treated with Vector, HSPA5-shRNA or Flag-HSPA5 plasmid. The CCK-8 assay kit was used to test the cell viability. The malondialdehyde detection kit was used to detect the level of malondialdehyde. Results: The cells showed the modest survival rate when the concentration of artesunate was 20 μmol/L. Flow cytometry showed the levels of lipid ROS and malondialdehyde in cells treated with artesunate increased(P<0.05), which could be reversed by deferoxamine; cell viability in the HSPA5-shRNA group treated with artesunate were lower than those in the Vector group(P<0.05). The level of malondialdehyde in the HSPA5shRNA group treated with artesunate were higher than those in the Vector group(P<0.05). Conclusion: Knockdown of HSPA5 may enhance the chemosensitivity of artesunate in human hepatic carcinoma SMMC7721 cells.