Three classes of abnormal cell differentiation are dysplasia, metaplasia, and anaplasia. Dysplasia indicates an abnormal arrangement of cells, usually arising from a disturbance in their normal growth behaviour. Some dysplasias are precursor lesions to cancer, whereas others are harmless and regress spontaneously. For example, dysplasia of the uterine cervix, called cervical intraepithelial neoplasia (CIN), may progress to cervical cancer. It can be detected by cervical smear cytology tests
Metaplasia is the conversion of one cell type into another. In fact, it is not usually the differentiated cells themselves that change but rather the stem cell population from which they are derived. Metaplasia commonly occurs where chronic tissue damage is followed by extensive regeneration. For example, squamous metaplasia of the bronchi occurs when the ciliated respiratory epithelial cells of people who smoke develop into squamous, or flattened, cells. In intestinal metaplasia of the stomach, patches resembling intestinal tissue arise in the gastric mucosa, often in association with gastric ulcers. Both of these types of metaplasia may progress to cancer.
Anaplasia is a loss of visible differentiation that can occur in advanced cancer. In general, early cancers resemble their tissue of origin and are described and classified by their pattern of differentiation. However, as they develop, they produce variants of more abnormal appearance and increased malignancy. Finally, a highly anaplastic growth can occur, in which the cancerous cells bear no visible relation to the parent tissue.
Differentiation from visibly undifferentiated precursor cells occurs during embryonic development, during metamorphosis of larval forms, and following the separation of parts in asexual reproduction. It also takes place in adult organisms during the renewal of tissues and the regeneration of missing parts. Thus, cell differentiation is an essential and ongoing process at all stages of life.
The visible differentiation of cells is only the last of a progressive sequence of states. In each state, the cell becomes increasingly committed toward one type of cell into which it can develop. States of commitment are sometimes described as “specification” to represent a reversible type of commitment and as “determination” to represent an irreversible commitment. Although states of specification and determination both represent differential gene activity, the properties of embryonic cells are not necessarily the same as those of fully differentiated cells. In particular, cells in specification states are usually not stable over prolonged periods of time.
Two mechanisms bring about altered commitments in the different regions of the early embryo: cytoplasmic localization and induction. Cytoplasmic localization is evident in the earliest stages of development of the embryo. During this time, the embryo divides without growth, undergoing cleavage divisions that produce separate cells called blastomeres. Each blastomere inherits a certain region of the original egg cytoplasm, which may contain one or more regulatory substances called cytoplasmic determinants. When the embryo has become a solid mass of blastomeres (called a morula), it generally consists of two or more differently committed cell populations—a result of the blastomeres having incorporated different cytoplasmic determinants. Cytoplasmic determinants may consist of mRNA or protein in a particular state of activation. An example of the influence of a cytoplasmic determinant is a receptor called Toll, located in the membranes of Drosophila (fruit fly) eggs. Activation of Toll ensures that the blastomeres will develop into ventral (underside) structures, while blastomeres containing inactive Toll will produce cells that will develop into dorsal (back) structures.
In induction, the second mechanism of commitment, a substance secreted by one group of cells alters the development of another group. In early development, induction is usually instructive; that is, the tissue assumes a different state of commitment in the presence of the signal than it would in the absence of the signal. Inductive signals often take the form of concentration gradients of substances that evoke a number of different responses at different concentrations. This leads to the formation of a sequence of groups of cells, each in a different state of specification. For example, in Xenopus (clawed frog) the early embryo contains a signaling centre called the organizer that secretes inhibitors of bone morphogenetic proteins (BMPs), leading to a ventral-to-dorsal (belly-to-back) gradient of BMP activity. The activity of BMP in the ventral region of the embryo suppresses the expression of transcription factors involved in the formation of the central nervous system and segmented muscles. Suppression ensures that these structures are formed only on the dorsal side, where there is decreased activity of BMP.
The final stage of differentiation often involves the formation of several types of differentiated cells from one precursor or stem cell population. Terminal differentiation occurs not only in embryonic development but also in many tissues in postnatal life. Control of this process depends on a system of lateral inhibition in which cells that are differentiating along a particular pathway send out signals that repress similar differentiation by their neighbours. For example, in the developing central nervous system of vertebrates, neurons arise from a simple tube of neuroepithelium, the cells of which possess a surface receptor called Notch. These cells also possess another cell surface molecule called Delta that can bind to and activate Notch on adjacent cells. Activation of Notch initiates a cascade of intracellular events that results in suppression of Delta production and suppression of neuronal differentiation. This means that the neuroepithelium generates only a few cells with high expression of Delta surrounded by a larger number of cells with low expression of Delta. High Delta production and low Notch activation makes the cells develop into neurons. Low Delta production and high Notch activation makes the cells remain as precursor cells or become glial (supporting) cells. A similar mechanism is known to produce the endocrine cells of the pancreas and the goblet cells of the intestinal epithelium. Such lateral inhibition systems work because cells in a population are never quite identical to begin with. There are always small differences, such as in the number of Delta molecules displayed on the cell surface. The mechanism of lateral inhibition amplifies these small differences, using them to bring about differential gene expression that leads to stable and persistent states of cell differentiation.
The biochemical basis of cell differentiation is the synthesis by the cell of a particular set of proteins, carbohydrates, and lipids. This synthesis is catalyzed by proteins called enzymes. Each enzyme in turn is synthesized in accordance with a particular gene, or sequence of nucleotides in the DNA of the cell nucleus. A particular state of differentiation, then, corresponds to the set of genes that is expressed and the level to which it is expressed.
Dolly the sheep; cloning Dolly the sheep was successfully cloned in 1996 by fusing the nucleus from a mammary-gland cell of a Finn Dorset ewe into an enucleated egg cell taken from a Scottish Blackface ewe. Carried to term in the womb of another Scottish Blackface ewe, Dolly was a genetic copy of the Finn Dorset ewe.Encyclopædia Britannica, Inc.
It is believed that all of an organism’s genes are present in each cell nucleus, no matter what the cell type, and that differences between tissues are not due to the presence or absence of certain genes but are due to the expression of some and the repression of others. In animals the best evidence for retention of the entire set of genes comes from whole animal cloning experiments in which the nucleus of a differentiated cell is substituted for the nucleus of a fertilized egg. In many species this can result in the development of a normal embryo that contains the full range of body parts and cell types. Likewise, in plants it is often possible to grow complete embryos from individual cells in tissue culture. Such experiments show that any nucleus has the genetic information required for the growth of a developing organism, and they strongly suggest that, for most tissues, cell differentiation arises from the regulation of genetic activity rather than the removal or destruction of unwanted genes. The only known exception to this rule comes from the immune system, where segments of DNA in developing white blood cells are slightly rearranged, producing a wide variety of antibody and receptor molecules.
At the molecular level there are many ways in which the expression of a gene can be differentially regulated in different cell types. There may be differences in the copying, or transcription, of the gene into RNA; in the processing of the initial RNA transcript into mRNA; in the control of mRNA movement to the cytoplasm; in the translation of mRNA to protein; or in the stability of mRNA. However, the control of transcription has the most influence over gene expression and has received the most detailed analysis.
The DNA in the cell nucleus exists in the form of chromatin, which is made up of DNA bound to histones (simple alkaline proteins) and other nonhistone proteins. Most of the DNA is complexed into repeating structures called nucleosomes, each of which contains eight molecules of histone. Active genes are found in parts of the DNA where the chromatin has an “open” configuration, in which regulatory proteins are able to gain access to the DNA. The degree to which the chromatin opens depends on chemical modifications of the outer parts of the histone molecules and on the presence or absence of particular nonhistone proteins. Transcriptional control is exerted with the help of regulatory sequences that are found associated with a gene, such as the promoter sequence, a region near the start of the gene, and enhancer sequences, regions that lie elsewhere within the DNA that augment the activity of enzymes involved in the process of transcription. Whether or not transcription occurs depends on the binding of transcription factors to these regulatory sequences. Transcription factors are proteins that usually possess a DNA-binding region, which recognizes the specific regulatory sequence in the DNA, and an effector region, which activates or inhibits transcription. Transcription factors often work by recruiting enzymes that add modifications (e.g., acetyl groups or methyl groups) to or remove modifications from the outer parts of the histone molecules. This controls the folding of the chromatin and the accessibility of the DNA to RNA polymerase and other transcription factors.
In general, it requires several transcription factors working in combination to activate a gene. For example, the chicken delta 1 crystallin gene, normally expressed only in the lens of the eye, has a promoter that contains binding sites for two activating transcription factors and an enhancer that contains binding sites for two other activating transcription factors. There is also an additional enhancer site that can bind either an activator (deltaEF3) or a repressor (deltaEF1). Successful transcription requires that all these sites are occupied by the correct transcription factors.
Fully differentiated cells are qualitatively different from one another. States of terminal differentiation are stable and persistent, both in the lifetime of the cell and in successive cell generations (in the case of differentiated types that are capable of continued cell division). The inherent stability of the differentiated state is maintained by various processes, including feedback activation of genes by their own products and repression of inactive genes. Chromatin structure may be important in maintaining states of differentiation, although it is still unclear whether this can be maintained during DNA replication, which involves temporary removal of chromosomal proteins and unwinding of the DNA double helix.
A type of differentiation control that is maintained during DNA replication is the methylation of DNA, which tends to recruit histone deacetylases and hence close up the structure of the chromatin. DNA methylation occurs when a methyl group is attached to the exterior, or sugar-phosphate side, of a cytosine (C) residue. Cytosine methylation occurs only on a C nucleotide when it is connected to a G (guanine) nucleotide on the same strand of DNA. These nucleotide pairings are called CG dinucleotides. One class of DNA methylase enzyme can introduce new methylations when required, whereas another class, called maintenance methylases, methylates CG dinucleotides in the DNA double helix only when the CG of the complementary strand is already methylated. Each time the methylated DNA is replicated, the old strand has the methyl groups and the new strand does not. The maintenance methylase will then add methyl groups to all the CGs opposite the existing methyl groups to restore a fully methylated double helix. This mechanism guarantees stability of the DNA methylation pattern, and hence the differentiated state, during the processes of DNA replication and cell division.
Adult organisms are composed of a number of distinct cell types. Cells are organized into tissues, each of which typically contains a small number of cell types and is devoted to a specific physiological function. For example, the epithelial tissue lining the small intestine contains columnar absorptive cells, mucus-secreting goblet cells, hormone-secreting endocrine cells, and enzyme-secreting Paneth cells. In addition, there exist undifferentiated dividing cells that lie in the crypts between the intestinal villi and serve to replace the other cell types when they become damaged or worn out. Another example of a differentiated tissue is the skeletal tissue of a long bone, which contains osteoblasts (large cells that synthesize bone) in the outer sheath and osteocytes (mature bone cells) and osteoclasts (multinucleate cells involved in bone remodeling) within the matrix.
In general, the simpler the overall organization of the animal, the fewer the number of distinct cell types that they possess. Mammals contain more than 200 different cell types, whereas simple invertebrate animals may have only a few different types. Plants are also made up of differentiated cells, but they are quite different from the cells of animals. For example, a leaf in a higher plant is covered with a cuticle layer of epidermal cells. Among these are pores composed of two specialized cells, which regulate gaseous exchange across the epidermis. Within the leaf is the mesophyll, a spongy tissue responsible for photosynthetic activity. There are also veins composed of xylem elements, which transport water up from the soil, and phloem elements, which transport products of photosynthesis to the storage organs.
The various cell types have traditionally been recognized and classified according to their appearance in the light microscope following the process of fixing, processing, sectioning, and staining tissues that is known as histology. Classical histology has been augmented by a variety of more discriminating techniques. Electron microscopy allows for higher magnifications. Histochemistry involves the use of coloured precipitating substrates to stain particular enzymes in situ. Immunohistochemistry uses specific antibodies to identify particular substances, usually proteins or carbohydrates, within cells. In situ hybridization involves the use of nucleic acid probes to visualize the location of specific messenger RNAs (mRNA). These modern methods have allowed the identification of more cell types than could be visualized by classical histology, particularly in the brain, the immune system, and among the hormone-secreting cells of the endocrine system.
Cancer can arise when the controlling factors over cell growth fail and allow a cell and its descendants to keep dividing at the expense of the organism. Studies of viruses that transform cultured cells and thus lead to the loss of control of cell growth have provided insight into the mechanisms that drive the formation of tumours. Transformed cells may differ from their normal progenitors by continuing to proliferate at very high densities, in the absence of growth factors, or in the absence of a solid substrate for support.
cancer-causing retrovirusesRetroviral insertion can convert a proto-oncogene, integral to the control of cell division, into an oncogene, the agent responsible for transforming a healthy cell into a cancer cell. An acutely transforming retrovirus (shown at top), which produces tumours within weeks of infection, incorporates genetic material from a host cell into its own genome upon infection, forming a viral oncogene. When the viral oncogene infects another cell, an enzyme called reverse transcriptase copies the single-stranded genetic material into double-stranded DNA, which is then integrated into the cellular genome. A slowly transforming retrovirus (shown at bottom), which requires months to elicit tumour growth, does not disrupt cellular function through the insertion of a viral oncogene. Rather, it carries a promoter gene that is integrated into the cellular genome of the host cell next to or within a proto-oncogene, allowing conversion of the proto-oncogene to an oncogene.Encyclopædia Britannica, Inc.
Major advances in the understanding of growth control have come from studies of the viral genes that cause transformation. These viral oncogenes have led to the identification of related cellular genes called protooncogenes. Protooncogenes can be altered by mutation or epigenetic modification, which converts them into oncogenes and leads to cell transformation. Specific oncogenes are activated in particular human cancers. For example, an oncogene called RAS is associated with many epithelial cancers, while another, called MYC, is associated with leukemias.
An interesting feature of oncogenes is that they may act at different levels corresponding to the multiple steps seen in the development of cancer. Some oncogenes immortalize cells so that they divide indefinitely, whereas normal cells die after a limited number of generations. Other oncogenes transform cells so that they grow in the absence of growth factors. A combination of these two functions leads to loss of proliferation control, whereas each of these functions on its own cannot. The mode of action of oncogenes also provides important clues to the nature of growth control and cancer. For example, some oncogenes are known to encode receptors for growth factors that may cause continuous proliferation in the absence of appropriate growth factors.
Loss of growth control has the added consequence that cells no longer repair their DNA effectively, and thus aberrant mitoses occur. As a result, additional mutations arise that subvert a cell’s normal constraints to remain in its tissue of origin. Epithelial tumour cells, for example, acquire the ability to cross the basal lamina and enter the bloodstream or lymphatic system, where they migrate to other parts of the body, a process called metastasis. When cells metastasize to distant tissues, the tumour is described as malignant, whereas prior to metastasis a tumour is described as benign.
Several studies have identified the transition from the G1 to the S phase as a crucial control point of the cell cycle. Stimuli are known to cause resting cells to proliferate by inducing them to leave G1 and begin DNA synthesis. These stimuli, called growth factors, are naturally occurring proteins specific to certain groups of cells in the body. They include nerve growth factor, epidermal growth factor, and platelet-derived growth factor. Such factors may have important roles in the healing of wounds as well as in the maintenance and growth of normal tissues. Many growth factors are known to act on the external membrane of the cell, by interacting with specialized protein receptor molecules. These respond by triggering further cellular changes, including an increase in calcium levels that makes the cell interior more alkaline and the addition of phosphate groups to the amino acid tyrosine in proteins. The complex response of cells to growth factors is of fundamental importance to the control of cell proliferation.
In prokaryotes, DNA synthesis can take place uninterrupted between cell divisions, and new cycles of DNA synthesis can begin before previous cycles have finished. In contrast, eukaryotes duplicate their DNA exactly once during a discrete period between cell divisions. This period is called the S (for synthetic) phase. It is preceded by a period called G1 (meaning “first gap”) and followed by a period called G2, during which nuclear DNA synthesis does not occur.
The four periods G1, S, G2, and M (for mitosis) make up the cell division cycle. The cell cycle characteristically lasts between 10 and 20 hours in rapidly proliferating adult cells, but it can be arrested for weeks or months in quiescent cells or for a lifetime in neurons of the brain. Prolonged arrest of this type usually occurs during the G1 phase and is sometimes referred to as G0. In contrast, some embryonic cells, such as those of fruit flies (vinegar flies), can complete entire cycles and divide in only 11 minutes. In these exceptional cases, G1 and G2 are undetectable, and mitosis alternates with DNA synthesis. In addition, the duration of the S phase varies dramatically. The fruit fly embryo takes only four minutes to replicate its DNA, compared with several hours in adult cells of the same species.
A specialized division of chromosomes called meiosis occurs during the formation of the reproductive cells, or gametes, of sexually reproducing organisms. Gametes such as ova, sperm, and pollen begin as germ cells, which, like other types of cells, have two copies of each gene in their nuclei. The chromosomes composed of these matching genes are called homologs. During DNA replication, each chromosome duplicates into two attached chromatids. The homologous chromosomes are then separated to opposite poles of the meiotic spindle by microtubules similar to those of the mitotic spindle. At this stage in the meiosis of germ cells, there is a crucial difference from the mitosis of other cells. In meiosis the two chromatids making up each chromosome remain together, so that whole chromosomes are separated from their homologous partners. Cell division then occurs, followed by a second division that resembles mitosis more closely in that it separates the two chromatids of each remaining chromosome. In this way, when meiosis is complete, each mature gamete receives only one copy of each gene instead of the two copies present in other cells.
In eukaryotes the processes of DNA replication and cell division occur at different times of the cell division cycle. During cell division, DNA condenses to form short, tightly coiled, rodlike chromosomes. Each chromosome then splits longitudinally, forming two identical chromatids. Each pair of chromatids is divided between the two daughter cells during mitosis, or division of the nucleus, a process in which the chromosomes are propelled by attachment to a bundle of microtubules called the mitotic spindle.
Mitosis can be divided into five phases. In prophase the mitotic spindle forms and the chromosomes condense. In prometaphase the nuclear envelope breaks down (in many but not all eukaryotes) and the chromosomes attach to the mitotic spindle. Both chromatids of each chromosome attach to the spindle at a specialized chromosomal region called the kinetochore. In metaphase the condensed chromosomes align in a plane across the equator of the mitotic spindle. Anaphase follows as the separated chromatids move abruptly toward opposite spindle poles. Finally, in telophase a new nuclear envelope forms around each set of unraveling chromatids.
An essential feature of mitosis is the attachment of the chromatids to opposite poles of the mitotic spindle. This ensures that each of the daughter cells will receive a complete set of chromosomes. The mitotic spindle is composed of microtubules, each of which is a tubular assembly of molecules of the protein tubulin (see above The cytoskeleton). Some microtubules extend from one spindle pole to the other, while a second class extends from one spindle pole to a chromatid. Microtubules can grow or shrink by the addition or removal of tubulin molecules. The shortening of spindle microtubules at anaphase propels attached chromatids to the spindle poles, where they unravel to form new nuclei.
The two poles of the mitotic spindle are occupied by centrosomes, which organize the microtubule arrays. In animal cells each centrosome contains a pair of cylindrical centrioles, which are themselves composed of complex arrays of microtubules. Centrioles duplicate at a precise time in the cell division cycle, usually close to the start of DNA replication.
After mitosis comes cytokinesis, the division of the cytoplasm. This is another process in which animal and plant cells differ. In animal cells cytokinesis is achieved through the constriction of the cell by a ring of contractile microfilaments consisting of actin and myosin, the proteins involved in muscle contraction and other forms of cell movement. In plant cells the cytoplasm is divided by the formation of a new cell wall, called the cell plate, between the two daughter cells. The cell plate arises from small Golgi-derived vesicles that coalesce in a plane across the equator of the late telophase spindle to form a disk-shaped structure. In this process, each vesicle contributes its membrane to the forming cell membranes and its matrix contents to the forming cell wall. A second set of vesicles extends the edge of the cell plate until it reaches and fuses with the sides of the parent cell, thereby completely separating the two new daughter cells. At this point, cellulose synthesis commences, and the cell plate becomes a primary cell wall
Before a cell can divide, it must accurately and completely duplicate the genetic information encoded in its DNA in order for its progeny cells to function and survive. This is a complex problem because of the great length of DNA molecules. Each human chromosome consists of a long double spiral, or helix, each strand of which consists of more than 100 million nucleotides
The duplication of DNA is called DNA replication, and it is initiated by complex enzymes called DNA polymerases. These progress along the molecule, reading the sequences of nucleotides that are linked together to make DNA chains. Each strand of the DNA double helix, therefore, acts as a template specifying the nucleotide structure of a new growing chain. After replication, each of the two daughter DNA double helices consists of one parental DNA strand wound around one newly synthesized DNA strand.
In order for DNA to replicate, the two strands must be unwound from each other. Enzymes called helicases unwind the two DNA strands, and additional proteins bind to the separated strands to stabilize them and prevent them from pairing again. In addition, a remarkable class of enzyme called DNA topoisomerase removes the helical twists by cutting either one or both strands and then resealing the cut. These enzymes can also untangle and unknot DNA when it is tightly coiled into a chromatin fibre.
In the circular DNA of prokaryotes, replication starts at a unique site called the origin of replication and then proceeds in both directions around the molecule until the two processes meet, producing two daughter molecules. In rapidly growing prokaryotes, a second round of replication can start before the first has finished. The situation in eukaryotes is more complicated, as replication moves more slowly than in prokaryotes. At 500 to 5,000 nucleotides per minute (versus 100,000 nucleotides per minute in prokaryotes), it would take a human chromosome about a month to replicate if started at a single site. Actually, replication begins at many sites on the long chromosomes of animals, plants, and fungi. Distances between adjacent initiation sites are not always the same; for example, they are closer in the rapidly dividing embryonic cells of frogs or flies than in adult cells of the same species.
Accurate DNA replication is crucial to ensure that daughter cells have exact copies of the genetic information for synthesizing proteins. Accuracy is achieved by a “proofreading” ability of the DNA polymerase itself. It can erase its own errors and then synthesize anew. There are also repair systems that correct genetic damage to DNA. For example, the incorporation of an incorrect nucleotide, or damage caused by mutagenic agents, can be corrected by cutting out a section of the daughter strand and recopying the parental strand.