Agrin promotes the proliferation, invasion and migration of rectal cancer cells via the WNT signaling pathway to contribute to rectal cancer progression.

Rectal cancer is the most common malignant tumor in the digestive system with rapidly metastasis and highly recurrence. Agrin (AGRN) is a proteoglycan involving in a large number of human cancers. However, how AGRN regulates the progression of rectal cancer remains largely unknown. We aimed to determine the biological role of AGRN and its mechanism in rectal cancer. AGRN expression in rectal cancer tissues was detected based on TCGA. The survival curve was plotted using the Kaplan-Meier method. qRT-PCR and western blot were utilized to examine the expression level of AGRN in cells. Cell proliferation, clonogenic ability, invasion, and migration of rectal cancer cells were analyzed by CCK-8, colony formation and transwell experiments. GSEA was employed for the analysis of the potential pathways-related with AGRN in rectal cancer. The activity of WNT pathway was determined by western blot. AGRN expression was dramatically increased in rectal cancer, and its up-regulation was associated with poorer prognosis of rectal cancer patients. AGRN expression was an independent factor for the prognosis of rectal cancer. AGRN inhibition suppressed rectal cancer cell growth, invasion, and migration, whereas AGRN overexpression facilitated these behaviors of rectal cancer cells in vitro. Mechanistically, WNT signaling pathway was enriched in high AGRN-expressing patients with rectal cancer. AGRN elevated the activity of WNT pathway through increasing Cyclin D1, C-Myc, p-GSK-3ß, and p-ß-catenin expression. Our present study indicated that AGRN might function as an oncogenic indicator in rectal cancer via activating the WNT pathway, which would help develop optimized therapeutic therapies for rectal cancer.

[IL-8 Induces Epithelial-to-mesenchymal Transition of Ovarian Carcinoma Cells: a Preliminary Study].

OBJECTIVE: To explore the effect of IL-8 on the epithelial-to-mesenchymal transition (EMT) in ovarian cancer,which will provide experimental basis for revealing related molecular mechanism in malignant metastasis of ovarian cancer. METHODS: The migration of ovarian cancer cell line SKOV3 cells was explored with Real time label free cell analysis (RTCA) after treatment with recombinant human IL-8.SKOV3 cells were co-cultured with IL-8 for 48 h,proteins involved in EMT were investigated via Western blot to explore the effect of IL-8 on the activation of the EMT. Invasion of SKOV3 cells after treatment with IL-8 were evaluated by transwell assay. RESULTS: According to the results of RTCA,after treatment with IL-8 for 48 h,the migration of SKOV3 cells was in platform phase. The treatment of IL-8 unregulated vimentin and snail and downregulated E-cadherin,which suggested that IL-8 induced EMT in ovarian cancer. The results of transwell test showed that invasive ability of IL-8 pretreated SKOV3 cells was enhanced (P<0.05). CONCLUSION: IL-8 can induce the EMT of ovarian cancer and enhance the invasion and migration of ovarian cancer.

[Efficacy of Le Fort operation and its impact on patients' quality of life].

OBJECTIVE: To evaluate the objective and subjective effects of Le Fort operation and its impact on patients' quality of life before and after operation. METHODS: A total of 48 patients with pelvic organ prolapse undergoing Le Fort operation from October 2005 to May 2010 were recruited. Their clinical data were retrospectively analyzed and regular follow-ups of pelvic examinations and questionnaires conducted. The questionnaires included pelvic floor distress inventory-short form 20 (PFDI-20) and pelvic floor distress impact questionnaire-short form 7 (PFIQ-7). RESULTS: Forty patients had complete follow-up data. Their mean follow-up period was 22.3 months (range: 12 - 42). Among them, there were recurrence (n = 1), new-onset stress urinary incontinence symptoms (n = 6) and new-onset colorectal-anal symptoms (n = 2). The postoperative scores of PFDI-20, PFIQ-7, UDI-6 and POPDI-6 decreased markedly (P < 0.05). CONCLUSION: Le Fort operation can significantly improve patients' quality of life and subjective prolapse symptoms and lower urinary tract. However its improvement of colorectal-anal symptoms remains to be defined.

[Survey of long term female pelvic floor function and sexual life status after total hysterectomy].

OBJECTIVE: To evaluate status of female pelvic floor function and sexual life after total hysterectomy. METHODS: From March 2001 to January 2004, 92 patients with undergoing hysterectomy due to benign gynecological diseases were enrolled in this study. They were followed up at outpatient department, including pelvic examination, filling in female sexual quality questionnaire, pelvic floor distress inventory short form 20 (PFDI-20), pelvic floor distress impact questionnaire short form 7 (PFIQ-7) and quality of sexual life of chinese women questionnaire. RESULTS: At 6 years after total hysterectomy, it was observed that 7 cases (7/92, 7.6%) were pelvic organ prolapse and 62 cases (62/92, 67.4%) were urinary incontinence. A median score of PFDI-20 were 4.67. A median score of PFIQ-7 were 0. Symptoms of pelvic floor dysfunction concentrated in lower urinary tract (58 cases with cough leak, 32 cases with spot urine leakage, 31 cases with frequent micturition, 24 cases with urgent urination)and bowel symptoms (26 cases with constipation, 24 cases with defecation urgency, 21 cases without fully drained stool). In the 68 patients filling in female sexual quality questionnaire, an average score were (127 ± 20) points. Female sexual quality questionnaire score, sexual satisfaction, sexual communication and adjustment, sexual response and sexual body image were positively correlated with the patients' income (r = 0.432, P = 0.007; r = 0.356, P = 0.028; r = 0.475, P = 0.003; r = 0.421, P = 0.009; r = 0.324, P = 0.047). CONCLUSIONS: Hysterectomy may have long-term effect on female pelvic floor function and sexual life. Quality of sexual life in those patients was positively correlated with income.

[Preliminary study on metabolism of collagen in uterus sacral ligament fibroblasts under stress].

OBJECTIVE: To detect the expression of type I collagen, type III collagen, prolyl-4-hydroxylases (PH4) and matrix metalloproteinase 1 (MMP-1) in sacral ligament fibroblasts under stress, to understand the collagen synthesis and metabolism in stress situations change. METHODS: Eight patients who underwent abdominal hysterectomy for uterine benign disease were enrolled in this study. Primary sacral ligament fibroblasts were isolated by explant. After mechanical loading, gene expression of type I, III collagen, PH4 and MMP-1 were measured. RESULTS: Stress of 8% continuing for 24 hours, collagen I (1.13 ± 0.24), collagen III (1.05 ± 0.31) mRNA expression and PH4 expression (1.11 ± 0.31) compared with static groups (1) showed increasing trends;when the stress were 4% and 12%, collagen I (0.86 ± 0.26 and 0.85 ± 0.25), collagen III showed increasing trends (0.74 ± 0.29 and 0.83 ± 0.38) mRNA expression were decreased. After removal of the stress, in the stress of 4% for 1 hour, collagen I (0.79 ± 0.40, 0.97 ± 0.24 and 1.46 ± 0.75), collagen III (0.86 ± 0.40, 0.99 ± 0.60 and 1.59 ± 0.82) and PH4 (1.11 ± 0.51, 1.17 ± 0.54 and 1.37 ± 0.39) mRNA expression increased gradually. In 8% stress group, collagen I mRNA expression (1.16 ± 0.62, 1.01 ± 0.51 and 1.05 ± 0.80) reached the peak in day 1, and collagen III (0.99 ± 0.69, 1.59 ± 0.55 and 1.03 ± 0.91) and PH4 (1.05 ± 0.31, 1.07 ± 0.80 and 0.85 ± 0.31) mRNA expression reached the peak in day 2, then decreased. 4% and 8% of the stress with time after the change, MMP-1 mRNA expression have peaked at day 1. CONCLUSIONS: Moderate stress could contribute to pelvic floor collagen synthesis, too much or too little stress is not conducive to the synthesis of collagen. Collagen I and collagen III on the stress response may be different, the former have faster reaction than the latter. PH4 were involved in the synthesis of collagen, while MMP-1 may play a role in collagen degradation.

[Clinical study on pubococcygeal line determined by dynamic magnetic resonance imaging used in evaluating pelvic organ prolapse].

OBJECTIVE: To investigate clinical value of pubococcygeal line (PCL) determined by dynamic magnetic resonance imaging (MRI) used in evaluating pelvic organ prolapse (POP), and investigate the relationship of pelvic organ prolapse quantitation (POP-Q) stage and pubococcygeal line (PCL) for the patient with POP. METHODS: Twenty patients with POP were evaluated by POP-Q stage and pelvic dynamic MRI examination simultaneously. Sagittal MRI images were acquired at rest and during maximal Valsalva using a fast gradient echo sequence two-dimensional fast low angle shot (FLASH) T(1) weighted image. The degree of prolapsed anterior vaginal wall, uterus and posterior vaginal wall were measured by PCL and compared with POP-Q system. RESULTS: There were 20 cases with cystocel diagnosed by POP-Q staging system, in which bladder neck or bladder base of 17 patients were under the PCL during maximum Valsalva. The concordance rate was 85% (17/20) between PCL and POP-Q stage. There were 19 cases with rectocele diagnosed by POP-Q, in which the anorectal junction of 4 patients' PCL descent below more than 2.5 cm. The concordance rate was 4/19 between PCL and POP-Q stage. There were 14 cases with uterine prolapse diagnosed by POP-Q staging system, in which uterine cervix of all descent below PCL. The concordance rate was 14/14 between PCL and POP-Q stage. However, it was noted that 5 cases did not reach POP-Q staging and their lowest uteri cervix were below or above PCL but less than 1 cm at maximal Valsalva. CONCLUSIONS: Compared with POP-Q staging system, the reference line of PCL determined by dynamic MRI could diagnose uterine prolapse accurately and anterior vaginal wall with greater clinical value, however it was limited in diagnosing posterior vaginal wall prolapse effectively. Therefore, the clinical value of PCL should be further studied for evaluating POP.

Status, challenges, and future prospects of stem cell therapy in pelvic floor disorders.

Pelvic floor disorders (PFDs) represent a group of common and frequently-occurring diseases that seriously affect the life quality of women, generally including stress urinary incontinence and pelvic organ prolapse. Surgery has been used as a treatment for PFD, but almost 30% of patients require subsequent surgery due to a high incidence of postoperative complications and high recurrence rates. Therefore, investigations of new therapeutic strategies are urgently needed. Stem cells possess strong multi-differentiation, self-renewal, immunomodulation, and angiogenesis abilities and they are able to differentiate into various cell types of pelvic floor tissues and thus provide a potential therapeutic approach for PFD. Recently, various studies using different autologous stem cells have achieved promising results by improving the pelvic ligament and muscle regeneration and conferring the tissue elasticity and strength to the damaged tissue in PFD, as well as reduced inflammatory reactions, collagen deposition, and foreign body reaction. However, with relatively high rates of complications such as bladder stone formation and wound infections, further studies are necessary to investigate the role of stem cells as maintainers of tissue homeostasis and modulators in early interventions including therapies using new stem cell sources, exosomes, and tissue-engineering combined with stem cell-based implants, among others. This review describes the types of stem cells and the possible interaction mechanisms in PFD treatment, with the hope of providing more promising stem cell treatment strategies for PFD in the future.