Heterocellular N-cadherin junctions enable nontransformed cells to inhibit the growth of adjacent transformed cells.
Heterocellular N-cadherin junctions enable nontransformed cells to inhibit the growth of adjacent transformed cells.
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DOI：
10.1186/s12964-021-00817-9
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发表时间：
2022-02-17
期刊：
影响因子：
--
通讯作者：
Goldberg GS
中科院分区：
文献类型：
--
作者：
Sheehan SA;Retzbach EP;Shen Y;Krishnan H;Goldberg GS
Src酪氨酸激酶磷酸化效应蛋白以诱导podoplanin（PDPN）受体的表达，从而促进肿瘤进展。然而，未转化的细胞可以正常化相邻转化细胞的生长和形态。转化的细胞必须逃避这个过程，称为“接触正常化”，成为侵入性和恶性。接触正常化需要转化细胞和非转化细胞之间的连接通讯。然而，介导这一过程的具体连接点尚未确定。本研究旨在鉴定接触正常化所需的连接蛋白。Src转化细胞和口腔鳞状细胞癌细胞与非转化细胞一起培养。通过荧光显微镜观察转化细胞和非转化细胞之间异细胞粘附连接的形成。CRISPR技术用于产生钙粘蛋白缺陷型和钙粘蛋白感受态非转化细胞，以确定接触正常化期间对粘附连接的需求。通过生长测定、免疫荧光、蛋白质印迹和RNA-seq分析与钙粘蛋白缺陷或钙粘蛋白感受态非转化细胞一起培养的转化细胞的接触归一化。此外，Src转化细胞表达PDPN下的组成型活性外源启动子被用来检查PDPN的能力，推翻接触正常化。我们发现，N-钙粘蛋白（N-Cdh）似乎介导接触正常化。表达N-Cdh的钙粘蛋白感受态细胞抑制培养中邻近转化细胞的生长，而钙粘蛋白缺陷细胞未能抑制这些细胞的生长。RNA-seq分析的结果表明，约10%的受接触正常化影响的转录本依赖于钙粘蛋白介导的通讯，这组基因包括PDPN。相反，钙粘蛋白缺陷细胞不能抑制PDPN表达或使相邻转化细胞的生长正常化。这些数据表明，非转化细胞形成异细胞钙粘蛋白连接，以抑制PDPN在相邻的转化细胞的表达。此外，我们发现，PDPN使转化细胞能够克服接触正常化的影响，在面对持续的N-Cdh表达。钙粘蛋白感受态细胞未能正常化表达PDPN的转化细胞的生长下的组成型活性外源启动子。非转化细胞与相邻的转化细胞形成钙粘蛋白连接，以降低PDPN表达，从而抑制肿瘤细胞增殖。当单个细胞获得使它们能够形成肿瘤的变化时，癌症就开始了。在癌症发展的这些开始阶段，正常细胞包围并直接接触癌细胞，以防止肿瘤形成并抑制癌症进展。这个过程称为接触规范化。癌细胞必须摆脱接触正常化才能发展成恶性癌症。接触正常化是一个广泛而强大的过程;然而，对这一过程中涉及的机制知之甚少。这项工作确定了在正常细胞和癌细胞之间形成接触所需的蛋白质，并探索了癌细胞超越接触正常化而发展为恶性癌症的途径。视频摘要在线版本包含补充材料，可通过10. 1186/s12964-021-00817-9获取。
The Src tyrosine kinase phosphorylates effector proteins to induce expression of the podoplanin (PDPN) receptor in order to promote tumor progression. However, nontransformed cells can normalize the growth and morphology of neighboring transformed cells. Transformed cells must escape this process, called “contact normalization”, to become invasive and malignant. Contact normalization requires junctional communication between transformed and nontransformed cells. However, specific junctions that mediate this process have not been defined. This study aimed to identify junctional proteins required for contact normalization. Src transformed cells and oral squamous cell carcinoma cells were cultured with nontransformed cells. Formation of heterocellular adherens junctions between transformed and nontransformed cells was visualized by fluorescent microscopy. CRISPR technology was used to produce cadherin deficient and cadherin competent nontransformed cells to determine the requirement for adherens junctions during contact normalization. Contact normalization of transformed cells cultured with cadherin deficient or cadherin competent nontransformed cells was analyzed by growth assays, immunofluorescence, western blotting, and RNA-seq. In addition, Src transformed cells expressing PDPN under a constitutively active exogenous promoter were used to examine the ability of PDPN to override contact normalization. We found that N-cadherin (N-Cdh) appeared to mediate contact normalization. Cadherin competent cells that expressed N-Cdh inhibited the growth of neighboring transformed cells in culture, while cadherin deficient cells failed to inhibit the growth of these cells. Results from RNA-seq analysis indicate that about 10% of the transcripts affected by contact normalization relied on cadherin mediated communication, and this set of genes includes PDPN. In contrast, cadherin deficient cells failed to inhibit PDPN expression or normalize the growth of adjacent transformed cells. These data indicate that nontransformed cells formed heterocellular cadherin junctions to inhibit PDPN expression in adjacent transformed cells. Moreover, we found that PDPN enabled transformed cells to override the effects of contact normalization in the face of continued N-Cdh expression. Cadherin competent cells failed to normalize the growth of transformed cells expressing PDPN under a constitutively active exogenous promoter. Nontransformed cells form cadherin junctions with adjacent transformed cells to decrease PDPN expression in order to inhibit tumor cell proliferation. Cancer begins when a single cell acquires changes that enables them to form tumors. During these beginning stages of cancer development, normal cells surround and directly contact the cancer cell to prevent tumor formation and inhibit cancer progression. This process is called contact normalization. Cancer cells must break free from contact normalization to progress into a malignant cancer. Contact normalization is a widespread and powerful process; however, not much is known about the mechanisms involved in this process. This work identifies proteins required to form contacts between normal cells and cancer cells, and explores pathways by which cancer cells override contact normalization to progress into malignant cancers. Video Abstract The online version contains supplementary material available at 10.1186/s12964-021-00817-9.
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DOI：
10.1002/gcc.20808
发表时间：
2010-11
期刊：
Genes, chromosomes & cancer
影响因子：
--
作者：
Cortez MA;Nicoloso MS;Shimizu M;Rossi S;Gopisetty G;Molina JR;Carlotti C Jr;Tirapelli D;Neder L;Brassesco MS;Scrideli CA;Tone LG;Georgescu MM;Zhang W;Puduvalli V;Calin GA
通讯作者：
Calin GA
影响因子：
5.7
作者：
Li, Xun;Jia, Zhenyu;Goldberg, Gary S.
通讯作者：
Goldberg, Gary S.
山槐种子凝集素（MASL）对口腔鳞状细胞癌（OSCC）基因表达和转录信号通路的影响。
影响因子：
3.6
作者：
Hamilton KL;Sheehan SA;Retzbach EP;Timmerman CA;Gianneschi GB;Tempera PJ;Balachandran P;Goldberg GS
通讯作者：
Goldberg GS
影响因子：
5
作者：
Bianconi D;Herac M;Spies D;Kieler M;Brettner R;Unseld M;Fürnkranz K;Famler B;Schmeidl M;Minichsdorfer C;Zielinski C;Heller G;Prager GW
通讯作者：
Prager GW
影响因子：
4.8
作者：
Fujii, Mizue;Honma, Masaru;Iizuka, Hajime
通讯作者：
Iizuka, Hajime