PD-L1 promotes colorectal cancer stem cell expansion by activating HMGA1-dependent signaling pathways.

PD-L1 is critical for tumor cell escape from immune surveillance by inhibiting T cell function via the PD-1 receptor. Accumulating evidence demonstrates that anti-PD-L1 monoclonal antibodies might potently enhance antitumor effects in various tumors, but the effect of PD-L1 on colorectal cancer stem cells (CSCs) remains unclear. We observed high PD-L1 expression in CD133+CD44+ colorectal CSCs and CSC-enriched tumorspheres. Altering PD-L1 expression promoted colorectal CSC self-renewal by increasing the expression of stemness genes, the CD133+CD44+ cell population sizes and the ability to form tumorspheres. Additionally, PD-L1 expression was markedly increased in chemoresistant colorectal cancer (CRC) cells in vitro and in vivo. More importantly, PD-L1 enhanced CRC cell tumorigenicity in nude mice; the inoculation of 1 × 104 cells resulted in high tumor formation efficiency. Mechanistically, PD-L1 directly interacted with HMGA1, and HMGA1 upregulation by PD-L1 activated HMGA1-dependent pathways, including the PI3K/Akt and MEK/ERK pathways, and promoted CSC expansion. HMGA1 downregulation rescued the PD-L1-induced phenotypes, highlighting the role of HMGA1 in PD-L1-mediated colorectal CSC self-renewal. Moreover, PD-L1 expression was correlated with the expression of CSC markers and HMGA1 in clinical CRC specimens. Thus, PD-L1 could crucially contribute to the maintenance of CSC self-renewal by activating HMGA1-dependent signaling pathways.

Long non­coding RNA 00152 functions as a competing endogenous RNA to regulate NRP1 expression by sponging with miRNA­206 in colorectal cancer.

Colorectal cancer (CRC) is the third most common type of cancer; however, the molecular mechanisms underlying colorectal tumor metastasis and growth remain elusive. Recently, accumulating evidence has indicated that long non­coding RNAs (lncRNAs) play a critical role in CRC progression and metastasis; however, the biological role and clinical significance of lncRNA 00152 (lnc00152) in CRC remains largely unknown. Thus, in this study, lnc00152 expression was measured in 80 human CRC tissue samples, 40 non­cancerous tissue samples, and 3 CRC cell lines (SW480, SW620 and LoVo) using RT­qPCR. We examined the effects of lnc00152 on CRC cells following transfection with lnc00152 overexpression plasmid or respective siRNA in vitro and in vivo. Luciferase assays revealed the mechanism driving competitive endogenous RNA (ceRNA). We identified that lnc00152 was aberrantly overexpressed in colorectal tumors and cancer cells and that lnc00152 was modulated by miRNA­206. lnc00152 overexpression enhanced the proliferative and invasive ability of CRC cells in vitro, promoted tumor growth in vivo, and was associated with the shorter overall survival of patients with CRC. In addition, lnc00152 overexpression promoted epithelial-mesenchymal transition (EMT) and increased neuropilin­1 (NRP1) expression in the CRC cells. By contrast, lnc00152 silencing exerted a counteractive effect. Collectively, these findings demonstrate the critical role of lnc00152 in tumor growth and progression in CRC, and identify a novel therapeutic target associated with CRC development and progression.

Gambogic Acid Efficiently Kills Stem-Like Colorectal Cancer Cells by Upregulating ZFP36 Expression.

BACKGROUND/AIMS: Gambogic acid (GA), the main active compound of Gamboge hanburyi, has been reported to be a potential novel antitumor drug. Whether GA inhibits putative cancer stem cells (CSCs), which are considered to be the major cause of cancer treatment failure, remains largely unknown. This study investigated whether GA inhibits the CSCs of colorectal cancer (CRC) and its possible mechanisms. METHODS: We performed CCK8 and tumor sphere formation assays, percentage analysis of both side population and CD133+CD44+ cells, and the detection of stem cells markers, in order to assess the role of GA in inhibiting the stem celllike features of CRC. An mRNA microarray was performed to identify the downstream gene affected by GA and rescue assays were performed to further clarify whether the downstream gene is involved in the GA induced decrease of the stem cell-like CRC population. CRC cells were engineered with a CSC detector vector encoding GFP and luciferase (Luc) under the control of the Nanog promoter, which were utilized to investigate the effect of GA on putative CSC in human tumor xenograft-bearing mice using in vivo bioluminescence imaging. RESULTS: Our results showed that GA significantly reduced tumor sphere formation and the percentages of side population and CD133+CD44+ cells, while also decreasing the expression of stemness and EMT-associated markers in CRC cells in vitro. GA killed stem-like CRC cells by upregulating the expression of ZFP36, which is dependent on the inactivation of the EGFR/ ERK signaling pathway. GFP+ cells harboring the PNanog-GFP-T2A-Luc transgene exhibited CSC characteristics. The in vivo results showed that GA significantly inhibited tumor growth in nude mice, accompanied by a remarkable reduction in the putative CSC number, based on whole-body bioluminescence imaging. CONCLUSION: These findings suggest that GA significantly inhibits putative CSCs of CRC both in vitro and in vivo by inhibiting the activation of the EGFR/ ERK/ZFP36 signaling pathway and may be an effective drug candidate for anticancer therapies.

Comparison of therapeutic effects of peripheral facial paralysis in acute stage by different interventions / 中国针灸

<p><b>OBJECTIVE</b>To compare the therapeutic effects of peripheral facial paralysis in acute stage by different interventions and explore the better treatments of peripheral facial paralysis.</p><p><b>METHODS</b>One hundred and thirty one cases of Bell's facial paralysis were randomly divided into three groups. In acupuncture group (44 cases), Dicang (ST 4), Jiache (ST 6), Hegu (LI 4), Yangbai (GB 14) and Taiyang (EX-HN 5), etc. were applied; in electroacupuncture group (45 cases), the selection of acupoints and needling method were same as those in acupuncture group, and the electroacupuncture therapy was applied on Dicang (ST 4), Xiaguan (ST 7), Yangbai (GB 14) and Taiyang (EX-HN 5) in acute stage; in medication and acupuncture group (42 cases), Prednisone and Acyclovir were taken by oral administration, Vitamin B1 and Vitamin B12, were applied by intramuscular injection in acute stage, and acupuncture was applied by the way which was same as that in acupuncture group during quiescent and recovery stages. The curative effects were evaluated by House-Brackmann Grading Scale, and the failed rates were observed by follow-up after one and three months.</p><p><b>RESULTS</b>The cured and markedly effective rates were 79.6% (35/44), 93.4% (42/45) and 78.6% (33/42) respectively in acupuncture group, electroacupuncture group and medication and acupuncture group, and the result in electroacupuncture group was superior to those in acupuncture group and medication and acupuncture group (P < 0.05). The cured rates above tympanichord were 54.2% (13/24), 85.2% (23/27) and 48.0% (12/25) in acupuncture group, electroacupuncture group and medication and acupuncture group, and the result in electroacupuncture group was superior to those in acupuncture group and medication and acupuncture group (P < 0.01). There was no significant differences of cured rates below tympanichord among three groups (P > 0.05); and the failed rate in electroacupuncture group was much lower than those in acupuncture group and medication and acupuncture group by follow-up after one and three months (all P < 0.01).</p><p><b>CONCLUSION</b>The peripheral facial paralysis is effectively treated by electroacupuncture in acute stage, and it suggests that electroacupuncture should be applied early during the acupuncture treatment of peripheral facial paralysis.</p>