Cldn4 overexpression promotes penile cavernous smooth muscle cell fibrotic response via the JNK signaling pathway.

BACKGROUND: Erectile dysfunction (ED), defined as the inability to achieve or maintain a penile erection sufficient to satisfy sexual behavior, is prevalent worldwide. AIM: Using previous research, bioinformatics, and experimental confirmation, we aimed to discover genes that contribute to ED through regulating hypoxia in corpus cavernosum smooth muscle cells (CCSMCs). METHODS: We used the Gene Expression Omnibus to acquire the sequencing data of the corpus cavernosum transcriptome for diabetic ED and nerve injury type ED rats. We intersected the common differentially expressed genes. Further verification was performed using single cell sequencing. Real-time quantitative polymerase chain reaction and immunofluorescence were used to investigate whether the differentially expressed genes are found in the corpus cavernosum. We used induced hypoxia to assess cell viability changes, and we developed a lentivirus overexpressing Cldn4 for in vitro and in vivo experiments to measure changes in JNK signaling, fibrosis, hypoxia, and erectile function. OUTCOMES: Our results indicate that targeting the JNK pathway and decreasing local hypoxia may be better options for therapeutic intervention to improve erectile function. RESULTS: We identified Cldn4 and found its expression increased in the corpora cavernosa of the 2 datasets. In addition, we found that hypoxia can increase the expression of Cldn4, activate the JNK signaling pathway, and exacerbate fibrosis in CCSMCs. Cldn4 overexpression in CCSMCs activated the JNK signaling pathway and increased fibrotic protein expression. Last, rat corpus cavernosum overexpressing Cldn4 activated the JNK signaling pathway, increased local fibrosis, and impaired erectile function. CLINICAL IMPLICATIONS: Through bioinformatics and in vitro and in vivo experiments, we found that Cldn4 has a negative effect on ED, and targeting Cldn4 may provide new ideas for ED treatment. STRENGTHS AND LIMITATIONS: Although we have identified Cldn4 as a potential target for ED treatment, we have only conducted preliminary validation on CCMSCs, and we still need to further validate in other cell lines. CONCLUSION: CCSMC hypoxia leads to increased Cldn4, in both nerve injury and diabetic ED rat models, and promotes fibrosis by activating the JNK signaling pathway.

Effect of umbilical cord blood dendritic cells induced by gastric cancer antigen combined with CIK cells in gastric cancer cell lines SGC-7901 / 国际肿瘤学杂志

Objective To investigate the effect of umbilical cord blood dendritic cells(DCs)induced by gastric cancer antigen combined with cytokine induced killer(CIK)cells in gastric cancer cell lines SGC-7901 in vitro. Methods Mononuclear cells from umbilical cord blood were used to create DCs and CIKs. The cell surface antigen expression of the mature DCs such as CD83,CD86,CD11c and the cell surface antigen of CIKs such as CD3,CD56,CD4,CD8,CD16 were detected using flow cytometry. Sensitized DCs-CIKs,DCs-CIKs,CIKs as effective cells,and SGC-7901 as target cells,the killing activities of these effective cells were tested with LDH release,which the number ratio of cells between effective cells and SGC-7901 cells were 10 :1,20 : 1,40 : 1,respectively. Results The cell surface antigen expressions of the mature DCs,such as CD83 + CD86 + ,CD11c + CD83 + ,CD86 + CD11c + were(75. 4 ± 2. 1)% ,(79. 3 ± 1. 4)% ,(80. 2 ± 2. 6)% , respectively. The mature sensitive-DCs surface antigen expressions,such as CD83 + CD86 + ,CD11c + CD83 + , CD86 + CD11c + ,were(77. 7 ± 1. 5)% ,(82. 6 ± 1. 9)% ,(76. 9 ± 2. 6)% ,respectively. There was no sta-tistical significance about the surface antigen expression between DCs and sensitive-DCs(t = 1. 526,P ﹥ 0. 05;t = 0. 958,P ﹥ 0. 05;t = 1. 049,P ﹥ 0. 05). The CIKs surface antigen expressions,such as CD4 + ,CD8 + , CD3 + CD56 + CD16 + ,were(22. 8 ± 1. 3)% ,(77. 3 ± 1. 8)% ,(24. 5 ± 2. 1)% ,respectively. The results suggested that the killing effect of the three kinds of combination cells on gastric cancer cells was different. The number ratio of cells between sensitive-DCs and SGC-7901 cells were 10 : 1,20 : 1,40 : 1,which the killing activities of sensitive-DCs-CIKs against SGC-7901 were(37. 68 ± 1. 49)% ,(41. 67 ± 0. 90)% ,(42. 71 ± 0. 98)% ,respectively. The killing activity of sensitive-DCs-CIKs was the highest when the ratio of cells between sensitive-DCs and SGC-7901 cells were 40 : 1. The killing activities of DC-CIKs were(36. 77 ± 0. 46)% ,(38. 94 ± 0. 95)% ,(41. 15 ± 0. 89)% ,respectively. The killing activities of CIKs were(34. 74 ± 1. 01)% ,(37. 76 ± 0. 43)% ,(39. 65 ± 0. 79)% ,respectively. There were statistically significant differences among the three groups(F = 5. 92,P ﹤ 0. 05;F = 19. 13,P ﹤ 0. 05;F = 8. 88,P ﹤ 0. 05). Conclusion The tumor killing activity of CIK is enhanced obviously by umbilical cord blood DCs which is sensitized by gastric cancer tumor antigen. There is the highest killing activity when the number ratio of cells between sensitive-DC-CIK and SGC-7901 cells is 40 : 1.