d6587bf8a8653e4a286614508c0c9a99 cancers_a2020v12n10p2798.pdf cd58fc03573c6347232da213056d3b1d49f69160 cancers_a2020v12n10p2798.pdf d75f86ee8f2526eb6db6b2779dd3561cb45cfbbda20f3aced0bfa9e0a7ecb386 cancers_a2020v12n10p2798.pdf Title: Cx43 and Associated Cell Signaling Pathways Regulate Tunneling Nanotubes in Breast Cancer Cells Subject: Connexin 43 (Cx43) forms gap junctions that mediate the direct intercellular diffusion of ions and small molecules between adjacent cells. Cx43 displays both pro- and anti-tumorigenic properties, but the mechanisms underlying these characteristics are not fully understood. Tunneling nanotubes (TNTs) are long and thin membrane projections that connect cells, facilitating the exchange of not only small molecules, but also larger proteins, organelles, bacteria, and viruses. Typically, TNTs exhibit increased formation under conditions of cellular stress and are more prominent in cancer cells, where they are generally thought to be pro-metastatic and to provide growth and survival advantages. Cx43 has been described in TNTs, where it is thought to regulate small molecule diffusion through gap junctions. Here, we developed a high-fidelity CRISPR/Cas9 system to knockout (KO) Cx43. We found that the loss of Cx43 expression was associated with significantly reduced TNT length and number in breast cancer cell lines. Notably, secreted factors present in conditioned medium stimulated TNTs more potently when derived from Cx43-expressing cells than from KO cells. Moreover, TNT formation was significantly induced by the inhibition of several key cancer signaling pathways that both regulate Cx43 and are regulated by Cx43, including RhoA kinase (ROCK), protein kinase A (PKA), focal adhesion kinase (FAK), and p38. Intriguingly, the drug-induced stimulation of TNTs was more potent in Cx43 KO cells than in wild-type (WT) cells. In conclusion, this work describes a novel non-canonical role for Cx43 in regulating TNTs, identifies key cancer signaling pathways that regulate TNTs in this setting, and provides mechanistic insight into a pro-tumorigenic role of Cx43 in cancer. Keywords: connexin 43; gap junctions; cancer; intercellular communication; breast cancer; cell signaling; tunneling nanotubes; cell–cell communication; tumor microtubes Author: Alexander Tishchenko, Daniel D. Azorín, Laia Vidal-Brime, María José Muñoz, Pol Jiménez Arenas, Christopher Pearce, Henrique Girao, Santiago Ramón y Cajal and Trond Aasen Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Tue Sep 29 13:07:13 2020 CEST ModDate: Tue Sep 29 13:07:13 2020 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 25 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4358806 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SNXGFU+URWPalladioL-Ital Type 1 Custom yes yes no 141 0 WYNAAJ+URWPalladioL-Bold Type 1 Custom yes yes no 142 0 CUAJJQ+Pxbsy Type 1 Builtin yes yes no 143 0 XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 144 0 AAHBMB+URWPalladioL-Roma Type 1 Custom yes yes no 145 0 YQFJFW+VnURWPalladioL Type 1 Custom yes yes no 146 0 SYXPPI+Rpxr Type 1 Builtin yes yes no 147 0 PUXWWS+Pxsy 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EMPGKH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 467 0 EMPGID+PalatinoLinotype TrueType WinAnsi yes yes no 495 0 EMPGKF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 496 0 EMPGKH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 497 0 EMPGID+PalatinoLinotype TrueType WinAnsi yes yes no 522 0 EMPGKF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 523 0 EMPGKH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 524 0 EMPGID+PalatinoLinotype TrueType WinAnsi yes yes no 555 0 EMPGKF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 556 0 EMPGKH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 557 0 EMPGIC+PalatinoLinotype CID TrueType Identity-H yes yes yes 596 0 EMPGID+PalatinoLinotype TrueType WinAnsi yes yes no 597 0 EMPGKF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 598 0 EMPGKH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 599 0 YICMPF+Rpcxr Type 1 Builtin yes yes no 678 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2020-11-26 06:09:48 CET RepresentationInformation: cancers_a2020v12n10p2798.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2020-11-11 13:11:53 CET Size: 4358806 Format: PDF Version: 1.5 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: ぉのぴひはつふっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸲ Item: Title: げづびふぬぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮ㄩ‾㸊⡜㌷㙜㌷㝜〰ぇおぁ〱ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰げづつふっづびぜ〴ぜ〰ごぎごぜ〴ぜ〰ぎふねぢづひぜ〴ぜ〰ちのつぜ〴ぜ〰がづのでぴとぜ〴ぜ〰どのぜ〴ぜ〰あご〵〴〹ぜ〴ぜ〰あひづちびぴぜ〴ぜ〰ぃちのっづひぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㈩‾㸊⡜㌷㙜㌷㝜〰ぇおぁ〱ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰げづつふっづびぜ〴ぜ〰ぴとづぜ〴ぜ〰ぎふねぢづひぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰えび〵〷〸ぴぜ〴ぜ〰あひづちびぴぜ〴ぜ〰ぃちのっづひぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰こどでのちぬどのでぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰がどのにづつぜ〴ぜ〰ぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰いどててづひづのぴどちぬぬべぜ〴ぜ〰ぁててづっぴぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴ぜ〰どのぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰しごぜ〴ぜ〰ちのつぜ〴ぜ〰かくぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰いひふで〭ぉのつふっづつぜ〴ぜ〰こぴどねふぬちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐひづびづのっづぜ〴ぜ〰はひぜ〴ぜ〰ぁぢびづのっづぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰どのぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰がどのづびぜ〴ぜ〰ぅへばひづびびどのでぜ〴ぜ〰ざづひべぜ〴ぜ〰えどでとぜ〴ぜ〰はひぜ〴ぜ〰ざづひべぜ〴ぜ〰がはぷぜ〴ぜ〰ぎふねぢづひびぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰ふのつづひぜ〴ぜ〰ぎはひねちぬぜ〴ぜ〰ぃはのつどぴどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㔩‾㸊⡜㌷㙜㌷㝜〰うふひぴとづひぜ〴ぜ〰ぉのびどでとぴぜ〴ぜ〰どのぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰こぴどねふぬちぴどのでぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸱ Children: Item: Title: ぇおぁ〱ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰げづつふっづびぜ〴ぜ〰ごぎごぜ〴ぜ〰ぎふねぢづひぜ〴ぜ〰ちのつぜ〴ぜ〰がづのでぴとぜ〴ぜ〰どのぜ〴ぜ〰あご〵〴〹ぜ〴ぜ〰あひづちびぴぜ〴ぜ〰ぃちのっづひぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㈩‾㸊⡜㌷㙜㌷㝜〰ぇおぁ〱ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰げづつふっづびぜ〴ぜ〰ぴとづぜ〴ぜ〰ぎふねぢづひぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰えび〵〷〸ぴぜ〴ぜ〰あひづちびぴぜ〴ぜ〰ぃちのっづひぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰こどでのちぬどのでぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰がどのにづつぜ〴ぜ〰ぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰いどててづひづのぴどちぬぬべぜ〴ぜ〰ぁててづっぴぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴ぜ〰どのぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰しごぜ〴ぜ〰ちのつぜ〴ぜ〰かくぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰いひふで〭ぉのつふっづつぜ〴ぜ〰こぴどねふぬちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐひづびづのっづぜ〴ぜ〰はひぜ〴ぜ〰ぁぢびづのっづぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰どのぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰がどのづびぜ〴ぜ〰ぅへばひづびびどのでぜ〴ぜ〰ざづひべぜ〴ぜ〰えどでとぜ〴ぜ〰はひぜ〴ぜ〰ざづひべぜ〴ぜ〰がはぷぜ〴ぜ〰ぎふねぢづひびぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰ふのつづひぜ〴ぜ〰ぎはひねちぬぜ〴ぜ〰ぃはのつどぴどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㔩‾㸊⡜㌷㙜㌷㝜〰うふひぴとづひぜ〴ぜ〰ぉのびどでとぴぜ〴ぜ〰どのぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰こぴどねふぬちぴどのでぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸱ Item: Title: ぇおぁ〱ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰げづつふっづびぜ〴ぜ〰ぴとづぜ〴ぜ〰ぎふねぢづひぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰えび〵〷〸ぴぜ〴ぜ〰あひづちびぴぜ〴ぜ〰ぃちのっづひぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰こどでのちぬどのでぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰がどのにづつぜ〴ぜ〰ぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰いどててづひづのぴどちぬぬべぜ〴ぜ〰ぁててづっぴぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴ぜ〰どのぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰しごぜ〴ぜ〰ちのつぜ〴ぜ〰かくぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰いひふで〭ぉのつふっづつぜ〴ぜ〰こぴどねふぬちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐひづびづのっづぜ〴ぜ〰はひぜ〴ぜ〰ぁぢびづのっづぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰どのぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰がどのづびぜ〴ぜ〰ぅへばひづびびどのでぜ〴ぜ〰ざづひべぜ〴ぜ〰えどでとぜ〴ぜ〰はひぜ〴ぜ〰ざづひべぜ〴ぜ〰がはぷぜ〴ぜ〰ぎふねぢづひびぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰ふのつづひぜ〴ぜ〰ぎはひねちぬぜ〴ぜ〰ぃはのつどぴどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㔩‾㸊⡜㌷㙜㌷㝜〰うふひぴとづひぜ〴ぜ〰ぉのびどでとぴぜ〴ぜ〰どのぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰こぴどねふぬちぴどのでぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸱ Item: Title: こどでのちぬどのでぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰がどのにづつぜ〴ぜ〰ぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰いどててづひづのぴどちぬぬべぜ〴ぜ〰ぁててづっぴぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴ぜ〰どのぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰しごぜ〴ぜ〰ちのつぜ〴ぜ〰かくぜ〴ぜ〰ぃづぬぬびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰いひふで〭ぉのつふっづつぜ〴ぜ〰こぴどねふぬちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐひづびづのっづぜ〴ぜ〰はひぜ〴ぜ〰ぁぢびづのっづぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰どのぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰がどのづびぜ〴ぜ〰ぅへばひづびびどのでぜ〴ぜ〰ざづひべぜ〴ぜ〰えどでとぜ〴ぜ〰はひぜ〴ぜ〰ざづひべぜ〴ぜ〰がはぷぜ〴ぜ〰ぎふねぢづひびぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰ふのつづひぜ〴ぜ〰ぎはひねちぬぜ〴ぜ〰ぃはのつどぴどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㔩‾㸊⡜㌷㙜㌷㝜〰うふひぴとづひぜ〴ぜ〰ぉのびどでとぴぜ〴ぜ〰どのぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰こぴどねふぬちぴどのでぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸱ Item: Title: いひふで〭ぉのつふっづつぜ〴ぜ〰こぴどねふぬちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰どのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐひづびづのっづぜ〴ぜ〰はひぜ〴ぜ〰ぁぢびづのっづぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰どのぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰がどのづびぜ〴ぜ〰ぅへばひづびびどのでぜ〴ぜ〰ざづひべぜ〴ぜ〰えどでとぜ〴ぜ〰はひぜ〴ぜ〰ざづひべぜ〴ぜ〰がはぷぜ〴ぜ〰ぎふねぢづひびぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰ふのつづひぜ〴ぜ〰ぎはひねちぬぜ〴ぜ〰ぃはのつどぴどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㔩‾㸊⡜㌷㙜㌷㝜〰うふひぴとづひぜ〴ぜ〰ぉのびどでとぴぜ〴ぜ〰どのぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰こぴどねふぬちぴどのでぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸱ Item: Title: うふひぴとづひぜ〴ぜ〰ぉのびどでとぴぜ〴ぜ〰どのぴはぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ぐちぴとぷちべびぜ〴ぜ〰こぴどねふぬちぴどのでぜ〴ぜ〰ごぎごぜ〴ぜ〰うはひねちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸱ Children: Item: Title: ごとづぜ〴ぜ〰ひはぬづぜ〴ぜ〰はてぜ〴ぜ〰ぇおぉぃぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸲ Item: Title: ぃとちののづぬ〭ぉのつづばづのつづのぴぜ〴ぜ〰ぅててづっぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸲⸵⸳ Item: Title: ぉのつふっぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ごぎごびぜ〴ぜ〰ぢべぜ〴ぜ〰ぃはのつどぴどはのづつぜ〴ぜ〰きづつどちぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: いどびっふびびどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸴ Item: Title: きちぴづひどちぬびぜ〴ぜ〰ちのつぜ〴ぜ〰きづぴとはつびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮ㄩ‾㸊⡜㌷㙜㌷㝜〰ぃへ〴〳ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰ちのつぜ〴ぜ〰くぶづひづへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ちのつぜ〴ぜ〰ぇおぁ〱〭〲〰にぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸱⸱ Children: Item: Title: ぃへ〴〳ぜ〴ぜ〰かのはっにはふぴぜ〴ぜ〰ちのつぜ〴ぜ〰くぶづひづへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ちのつぜ〴ぜ〰ぇおぁ〱〭〲〰にぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸱⸱ Children: Item: Title: ぇづのづひちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰ちのぜ〴ぜ〰ぁぬぬ〭どの〭くのづぜ〴ぜ〰えどでと〭うどつづぬどぴべぜ〴ぜ〰ぃぢと〭とてぃちび〹〭〲ぁ〭づぇうぐぜ〴ぜ〰ぃげぉこぐげぜ〴ぜ〰ぐぬちびねどつぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸱⸲ Item: Title: ぃぢと〭とてぃちび〹〭〲ぁ〭づぇうぐぜ〴ぜ〰ぃげぉこぐげぜ〴ぜ〰こべびぴづねぜ〴ぜ〰ごちひでづぴどのでぜ〴ぜ〰ぃへ〴〳ぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸱⸳ Item: Title: ぇづのづひちぴどはのぜ〴ぜ〰はてぜ〴ぜ〰かくぜ〴ぜ〰ぃぬはのづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸱⸴ Item: Title: くぶづひづへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぃへ〴〳ぜ〴ぜ〰ちのつぜ〴ぜ〰ぇおぁ〱〭〲〰にぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㈩‾㸊⡜㌷㙜㌷㝜〰ぃづぬぬぜ〴ぜ〰ぃふぬぴふひづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰ごぎごぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰いひふでぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぐづばぴどつづぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぃはのつどぴどはのづつぜ〴ぜ〰きづつどふねぜ〴ぜ〰ごひづちぴねづのぴぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぃづぬぬぜ〴ぜ〰ぃふぬぴふひづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰ごぎごぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰いひふでぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぐづばぴどつづぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぃはのつどぴどはのづつぜ〴ぜ〰きづつどふねぜ〴ぜ〰ごひづちぴねづのぴぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ごぎごぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰いひふでぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぐづばぴどつづぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぃはのつどぴどはのづつぜ〴ぜ〰きづつどふねぜ〴ぜ〰ごひづちぴねづのぴぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: いひふでぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰ぐづばぴどつづぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぃはのつどぴどはのづつぜ〴ぜ〰きづつどふねぜ〴ぜ〰ごひづちぴねづのぴぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぐづばぴどつづぜ〴ぜ〰ごひづちぴねづのぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ぃはのつどぴどはのづつぜ〴ぜ〰きづつどふねぜ〴ぜ〰ごひづちぴねづのぴぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぃはのつどぴどはのづつぜ〴ぜ〰きづつどふねぜ〴ぜ〰ごひづちぴねづのぴぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぐひはぴづどのぜ〴ぜ〰ぅへぴひちっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどてどっちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: しづびぴづひのぜ〴ぜ〰あぬはぴぴどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㤩‾㸊⡜㌷㙜㌷㝜〰ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: ぉねねふのはてぬふはひづびっづのっづぜ〴ぜ〰きどっひはびっはばべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㄰ Item: Title: う〭ちっぴどの〯ぇ〭ちっぴどのぜ〴ぜ〰げちぴどはぜ〴ぜ〰いづぴづひねどのちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮ㄱ Item: Title: ぐちひちっとふぴづぜ〴ぜ〰ぁびびちべぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮ㄲ Item: Title: こぴちぴどびぴどっちぬぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸵ Item: Title: ぃはのっぬふびどはのびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸵ Item: Title: げづてづひづのっづび Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: XWOCXD+VnURWPalladioL-Bold FontSubset: true FirstChar: 178 LastChar: 226 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